72 results on '"Guzzo, C."'
Search Results
2. Pilot Study of a Novel Therapeutic Approach for Refractory Advanced Stage Folliculotropic Mycosis Fungoides
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Guzzo, C, primary, Jariwala, N, additional, Haun, P, additional, MacArthur, K, additional, Mowad, C, additional, Richard, E, additional, Holton, J, additional, and Rook, A, additional
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- 2020
- Full Text
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3. Infliximab Inhibits Progression of Radiographic Damage in Patients With Active Psoriatic Arthritis Through One Year of Treatment: Results From the Induction and Maintenance Psoriatic Arthritis Clinical Trial 2
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van der Heijde, D., Kavanaugh, A., Gladman, D. D., Antoni, C., Krueger, G. G., Guzzo, C., Zhou, B., Dooley, L. T., de Vlam, K., Geusens, P., Birbara, C., Halter, D., and Beutler, A.
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- 2007
4. Functional Status Improvement In Infliximab-Treated Psoriatic Arthritis Patients Regardless of Baseline Radiographic Damage: IMPACT 2
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Kavanaugh, A, Krueger, G G, de Vlam, K, Birbara, C, Beutler, A, Dooley, L T, Guzzo, C, Bala, M, Kirkham, B, and Antoni, C
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- 2006
5. Infliximab improves signs and symptoms of psoriatic arthritis: results of the IMPACT 2 trial
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Antoni, C, Krueger, G, de Vlam, K, Birbara, C, Beutler, A, Guzzo, C, Zhou, B, Dooley, L, Kavanaugh, A, and t for
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Adult ,Male ,musculoskeletal diseases ,medicine.medical_specialty ,Immunology ,Arthritis ,Placebo ,Severity of Illness Index ,Gastroenterology ,General Biochemistry, Genetics and Molecular Biology ,Dactylitis ,Psoriatic arthritis ,Double-Blind Method ,Rheumatology ,Psoriasis Area and Severity Index ,Psoriasis ,Internal medicine ,medicine ,Humans ,Immunology and Allergy ,Tumor Necrosis Factor-alpha ,business.industry ,Arthritis, Psoriatic ,Therapeutic effect ,Antibodies, Monoclonal ,Middle Aged ,medicine.disease ,Infliximab ,Surgery ,Extended Report ,Treatment Outcome ,Antirheumatic Agents ,Female ,business ,medicine.drug - Abstract
Objectives: To evaluate further in a phase III, double blind trial the efficacy of infliximab in patients with active psoriatic arthritis (PsA), as observed in the smaller IMPACT trial. Methods: 200 patients with active PsA unresponsive to previous treatment were randomised to infusions of infliximab 5 mg/kg or placebo at weeks 0, 2, 6, 14, and 22. Patients with inadequate response entered early escape at week 16. The primary measure of clinical response was ACR20. Other measures included Psoriatic Arthritis Response Criteria (PsARC), Psoriasis Area and Severity Index (PASI), and dactylitis and enthesopathy assessments. Results: At week 14, 58% of patients receiving infliximab and 11% of those receiving placebo achieved an ACR20 response and 77% of infliximab patients and 27% of placebo patients achieved PsARC (both p
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- 2005
6. Stromal Cell-Derived Factor 2: A Novel Protein that Interferes in Endoplasmic Reticulum Stress Pathway in Human Placental Cells
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Lorenzon-Ojea, A. R., primary, Guzzo, C. R., additional, Kapidzic, M., additional, Fisher, S. J., additional, and Bevilacqua, E., additional
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- 2016
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7. 133 The incidence of cutaneous T-Cell lymphoma in the Veteran population
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Del Guzzo, C., primary, Levin, A., additional, Dana, A., additional, Vinnakota, R., additional, Park, Y., additional, Newman, J., additional, Langhoff, E., additional, and Geskin, L., additional
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- 2016
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8. Ustekinumab induction and maintenance therapy in refractory Crohn's disease
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Sandborn, Wj, Gasink, C, Gao, Ll, Blank, Ma, Johanns, J, Guzzo, C, Sands, Be, Hanauer, Sb, Targan, S, Rutgeerts, P, Ghosh, S, de Villiers WJ, Panaccione, R, Greenberg, G, Schreiber, S, Lichtiger, S, Feagan, Bg, Haas, T, Kaser, A, Vogelsang, H, Brown, S, Florin, T, Gibson, Pg, Hetzel, D, Leong, R, Pavli, P, Radford-Smith, G, Sparrow, M, Baert, F, D'Haens, G, D'Heygere, F, Franchimont, D, Louis, E, Mana, F, Moreels, T, Vermeire, S, Anderson, F, Axler, J, Greenbloom, S, Bitton, A, Fedorak, Rn, Larkai, E, Marshall, J, Singh, R, Abitbol, V, Allez, M, Lemann, M, Bonaz, B, Colombel, Jf, Dupas, Jl, Hebuterne, X, Laharie, D, Lerebours, E, Moreau, J, Bokemeyer, B, Holler, B, Howaldt, Sp, Krummenerl, T, Kucharzik, T, Ochsenkühn, T, Raedler, A, Schiefke, I, Seidler, U, Sturm, A, Zeitz, M, Eliakim, A, Fishman, S, Konikoff, Fm, Lavy, A, Niv, Y, Nussinson, E, Rachmilewitz, D, Andriulli, A, Annese, V, Biancone, L, Corazza, Gr, Danese, S, Sturniolo, Gc, Terrosu, G, Sorrentino, D, Hommes, Dw, Jansen, Jm, Otten, Mh, Pierik, M, Ponsioen, Cy, Stokkers, P, van Bodegraven AA, Van der Woude, J, Calvet Calvo, X, Casellas, F, Garcia López, S, Garcia-Planella, E, Ginard-Vincens, D, López San Román, A, Muñoz Nuñez, F, Pérez Gisbert, J, Vera, Mi, Rodrigo, J, Riestra-Menendez, S, Arnott, I, Bloom, S, Campbell, Ss, Harbord, Mw, Mansfield, Jc, Nwokolo, C, Parkes, M, Probert, Cs, Aberra, Fn, Abraham, Bp, Abreu, Mt, Amontree, Js, Barish, Cf, Barto, Ae, Behm, B, Birbara, Ca, Bologna, S, Dryden GW Jr, Eisner, Ms, Ertan, A, Fogel, R, Gagneja, Hk, Ginsburg, P, Goff, Js, Gordon, G, Hamilton, Jw, Hanson, Js, Hardi, R, Hemaidan, A, Higgins, P, Holderman, W, Hornbuckle, K, Ibegbu, E, Isaacs, Kl, Katz, Ja, Katz, S, Kaufman, Bp, Kavanaugh, Af, Khurana, Sk, Lashner, B, Lawrence, S, Hansen, Rn, Lee, S, Leighton, Ja, Leman, Bi, Levenson, Sd, Lowe, Je, Marcuard, Sp, Matsuyama, Rm, Mcnair, Ae, Melmed, G, Miller, Km, Miner PB Jr, Mutlu, Ea, Keshavarzian, A, Narayen, V, Noar, Md, Patel, Ph, Patrick, Tj, Peck, A, Peterson, Ka, Phillips, Rw, Picco, Mf, Randall, C, Richards, Rj, Safdi, Ma, Scherl, Eh, Schwartz, Da, Schwartz, Hi, Schwartz, Jl, Sedghi, S, Shafran, I, Siegel, Ca, Sninsky, Ca, Stern, M, Suiter, D, Swaminath, A, Terdiman, Jp, Mahadevan, U, Thomson, C, Valentine, J, Vasudeva, R, Vecchio, Ja, Wolf, Dc, Yajnik, V, Yabkowski, J, Yen, E., AGEM - Amsterdam Gastroenterology Endocrinology Metabolism, Gastroenterology and Hepatology, and Other departments
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Male ,Oncology ,MONOCLONAL-ANTIBODY ,Drug Resistance ,Disease ,Severity of Illness Index ,Crohn Disease ,Maintenance therapy ,IL-23 ,Adult ,Antibodies, Monoclonal ,Antibodies, Monoclonal, Humanized ,Double-Blind Method ,Female ,Humans ,Induction Chemotherapy ,Maintenance Chemotherapy ,Middle Aged ,Remission Induction ,Tumor Necrosis Factors ,Ustekinumab ,Monoclonal ,Humanized ,Settore MED/12 - Gastroenterologia ,Crohn's disease ,EXPERIMENTAL COLITIS ,General Medicine ,medicine.drug ,medicine.medical_specialty ,MONOCLONAL-ANTIBODY, RANDOMIZED-TRIAL, EXPERIMENTAL COLITIS, CERTOLIZUMAB PEGOL, INFLAMMATION, IL-23, INTERLEUKIN-12, ADALIMUMAB, INFLIXIMAB, DISCOVERY ,Antibodies ,CERTOLIZUMAB PEGOL ,INFLAMMATION ,Refractory ,Internal medicine ,INFLIXIMAB ,medicine ,business.industry ,Induction chemotherapy ,medicine.disease ,RANDOMIZED-TRIAL ,INTERLEUKIN-12 ,Clinical trial ,DISCOVERY ,Immunology ,Tumor Necrosis Factor Inhibitors ,business ,ADALIMUMAB - Abstract
BACKGROUND In patients with Crohn's disease, the efficacy of ustekinumab, a human monoclonal antibody against interleukin-12 and interleukin-23, is unknown. METHODS We evaluated ustekinumab in adults with moderate-to-severe Crohn's disease that was resistant to anti-tumor necrosis factor (TNF) treatment. During induction, 526 patients were randomly assigned to receive intravenous ustekinumab (at a dose of 1, 3, or 6 mg per kilogram of body weight) or placebo at week 0. During the maintenance phase, 145 patients who had a response to ustekinumab at 6 weeks underwent a second randomization to receive subcutaneous injections of ustekinumab (90 mg) or placebo at weeks 8 and 16. The primary end point was a clinical response at 6 weeks. RESULTS The proportions of patients who reached the primary end point were 36.6%, 34.1%, and 39.7% for 1, 3, and 6 mg of ustekinumab per kilogram, respectively, as compared with 23.5% for placebo (P = 0.005 for the comparison with the 6-mg group). The rate of clinical remission with the 6-mg dose did not differ significantly from the rate with placebo at 6 weeks. Maintenance therapy with ustekinumab, as compared with placebo, resulted in significantly increased rates of clinical remission (41.7% vs. 27.4%, P = 0.03) and response (69.4% vs. 42.5%, P < 0.001) at 22 weeks. Serious infections occurred in 7 patients (6 receiving ustekinumab) during induction and 11 patients (4 receiving ustekinumab) during maintenance. Basal-cell carcinoma developed in 1 patient receiving ustekinumab. CONCLUSIONS Patients with moderate-to-severe Crohn's disease that was resistant to TNF antagonists had an increased rate of response to induction with ustekinumab, as compared with placebo. Patients with an initial response to ustekinumab had significantly increased rates of response and remission with ustekinumab as maintenance therapy. (Funded by Janssen Research and Development; CERTIFI ClinicalTrials.gov number, NCT00771667.)
- Published
- 2012
9. Il carcinoma della mammella
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Guzzo, C, Lintas, F, Gueli, Nicolo', and Marigliano, Vincenzo
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- 2007
10. Chronic urticaria and blastocystis hominis infection: a case report
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Pasqui, A. L., Savini, E., Saletti, M., Guzzo, C., LUCA PUCCETTI, and Auteri, A.
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Urticaria ,Gastrointestinal Diseases ,Paromomycin ,Blastocystis Infections ,Middle Aged ,Drug Administration Schedule ,Diagnosis, Differential ,Drug Hypersensitivity ,Feces ,Italy ,Metronidazole ,Chronic Disease ,Animals ,Humans ,Blastocystis hominis ,Female ,Food Hypersensitivity - Abstract
We report a case of a 45 year old woman which fulfilled the criteria of chronic urticaria (remitting and relapsing bouts of erythematous and pruriginuos lesions without angioedema, lasted four months). Cutaneous manifestations were not related to a specific inducing factor, had no benefit from antihystamine and steroid drugs and were associated sometimes with mild gastroentric disorders. Patient was submitted to extensive clinical, laboratory and intrumental investigations which permit to exclude many conditions: allergy to inhalants, food, insects and drug adverse reactions, autoimmune urticaria, autoimmune diseases, neoplastic and infectious diseases. Finally coprocolture disclosed the presence of Blastocystis hominis in stool samples thus permitting to associate urticaria to parasitic infection. Both cutaneous manifestations and mild abdomen disturbs disappeared after appropriate treatment. Despite the high diffusion the aetiopathogenesis of chronic urticaria remains often undefined. A large number of parasites have been correlated with urticaria but few data exist as regards Blastocystis hominis infection; then our findings may add evidence to the role of this parasite in inducing chronic urticaria. Considering that Blastocystis hominis is a modest pathogen for humans, the mechanism is probably the typical one of cutaneous allergic hypersensitivity; antigen parasites induce the activation of specific clones of Th2 lymphocytes, the release of related cytokines and the consequent IgE production.
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- 2004
11. Comparison of ustekinumab and etanercept for moderate-to-severe psoriasis.
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Griffiths, C.E., Strober, B.E., Kerkhof, P.C.M. van de, Ho, V., Fidelus-Gort, R., Yeilding, N., Guzzo, C., Xia, Y., Zhou, B., Li, S., Dooley, L.T., Goldstein, N.H., Menter, A., Griffiths, C.E., Strober, B.E., Kerkhof, P.C.M. van de, Ho, V., Fidelus-Gort, R., Yeilding, N., Guzzo, C., Xia, Y., Zhou, B., Li, S., Dooley, L.T., Goldstein, N.H., and Menter, A.
- Abstract
Contains fulltext : 89832.pdf (publisher's version ) (Closed access), BACKGROUND: Biologic agents offer a range of new therapeutic options for patients with psoriasis; however, the relative benefit-risk profiles of such therapies are not well known. We compared two biologic agents, ustekinumab (an interleukin-12 and interleukin-23 blocker) and etanercept (an inhibitor of tumor necrosis factor alpha), for the treatment of psoriasis. METHODS: We randomly assigned 903 patients with moderate-to-severe psoriasis to receive subcutaneous injections of either 45 or 90 mg of ustekinumab (at weeks 0 and 4) or high-dose etanercept (50 mg twice weekly for 12 weeks). The primary end point was the proportion of patients with at least 75% improvement in the psoriasis area-and-severity index (PASI) at week 12; a secondary end point was the proportion with cleared or minimal disease on the basis of the physician's global assessment. Assessors were unaware of the treatment assignments. The efficacy and safety of a crossover from etanercept to ustekinumab were evaluated after week 12. RESULTS: There was at least 75% improvement in the PASI at week 12 in 67.5% of patients who received 45 mg of ustekinumab and 73.8% of patients who received 90 mg, as compared with 56.8% of those who received etanercept (P=0.01 and P<0.001, respectively). Similarly, 65.1% of patients who received 45 mg of ustekinumab and 70.6% of patients who received 90 mg of ustekinumab had cleared or minimal disease according to the physician's global assessment, as compared with 49.0% of those who received etanercept (P<0.001 for both comparisons). Among patients who did not have a response to etanercept, 48.9% had at least 75% improvement in the PASI within 12 weeks after crossover to ustekinumab. One or more adverse events occurred through week 12 in 66.0% of patients who received 45 mg of ustekinumab and 69.2% of patients who received 90 mg of ustekinumab and in 70.0% who received etanercept; 1.9%, 1.2%, and 1.2%, respectively, had serious adverse events. Safety patterns were simila
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- 2010
12. Nuevas herramientas de fertilización: impacto de la nanotecnología en el desarrollo temprano de Curcubita maxima.
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Domínguez, R. E., Brugo Carivali, F., Giachero, M. L., Guzzo, C., Perotto, C., and Ciacci, M. B.
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- 2022
13. Studio sull’associazione tra lipoproteina (a) e retinoparia aterosclerotica documentata mediante fluoroangiografia
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Montagnani, M, Motolese, Eduardo, Mulinari, M, Montomoli, M, Addabbo, G, Fiorentini, P, Guzzo, C, Di Luzio, M, Montagnani, A, and Gennari, C.
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- 1994
14. PSS44 USTEKINUMAB IMPROVES WORK PRODUCTIVITY AND DECREASES WORKDAYS MISSED DUE TO PSORIASIS IN PATIENTS WITH MODERATE TO SEVERE PSORIASIS
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Reich, K, primary, Lebwohl, M, additional, Schenkel, B, additional, Eisenberg, D, additional, Szapary, P, additional, Yeilding, N, additional, Guzzo, C, additional, Hsu, MC, additional, Li, S, additional, and Gordon, KB, additional
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- 2008
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15. PSS29 USTEKINUMAB IMPROVES DISEASE SPECIFIC HEALTH-RELATED QUALITY OF LIFE IN PATIENTS WITH MODERATE TO SEVERE PSORIASIS: RESULTS WITH THE DERMATOLOGY LIFE QUALITY INDEX
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Lebwohl, M, primary, Papp, K, additional, Schenkel, B, additional, Eisenberg, D, additional, Yeilding, N, additional, Guzzo, C, additional, Wang, Y, additional, Li, S, additional, and Krueger, GG, additional
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- 2008
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16. PSS30 USTEKINUMAB SIGNIFICANTLY IMPROVES QUALITY OF LIFE IN PATIENTS WITH PSORIASIS: RESULTS FROM A PHASE III STUDY
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Langley, R, primary, Lebwohl, M, additional, Krueger, GG, additional, Yeilding, N, additional, Guzzo, C, additional, Wang, Y, additional, Li, S, additional, Schenkel, B, additional, Reich, K, additional, and Leonardi, C, additional
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- 2008
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17. Clinical and Cognitive Phenotype of Mild Cognitive Impairment Evolving to Dementia with Lewy Bodies
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Simona Gardini, Caterina Guzzo, Mario Ermani, Francesca Gnoato, Cinzia Bussè, Micaela Mitolo, Annachiara Cagnin, Paolo Caffarra, Nela Jelcic, Cagnin A., Busse C., Gardini S., Jelcic N., Guzzo C., Gnoato F., Mitolo M., Ermani M., and Caffarra P.
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medicine.medical_specialty ,Dementia with Lewy bodie ,genetic structures ,Cognitive Neuroscience ,Dementia with Lewy bodies ,lcsh:Geriatrics ,Audiology ,behavioral disciplines and activities ,REM sleep behavior disorder ,lcsh:RC346-429 ,Rating scale ,mental disorders ,medicine ,Original Research Article ,Psychiatry ,Episodic memory ,lcsh:Neurology. Diseases of the nervous system ,Mini–Mental State Examination ,medicine.diagnostic_test ,Mini Mental State Examination ,Mild cognitive impairment ,Cognition ,medicine.disease ,Executive functions ,nervous system diseases ,lcsh:RC952-954.6 ,Psychiatry and Mental health ,Visuoconstructional abilities ,Psychology ,Neuropsychiatric Inventory Questionnaire - Abstract
Objective: The aim of this study was to determine which characteristics could better distinguish dementia with Lewy bodies (DLB) from Alzheimer's disease (AD) at the mild cognitive impairment (MCI) stage, with particular emphasis on visual space and object perception abilities. Methods: Fifty-three patients with mild cognitive deficits that were eventually diagnosed with probable DLB (MCI-DLB: n = 25) and AD (MCI-AD: n = 28) at a 3-year follow-up were retrospectively studied. At the first visit, the patients underwent cognitive assessment including the Qualitative Scoring Mini Mental State Examination Pentagon Test and the Visual Object and Space Perception Battery. The Neuropsychiatric Inventory Questionnaire, Unified Parkinson's Disease Rating Scale (UPDRS) and questionnaires for cognitive fluctuations and sleep disorders were also administered. Results: The best clinical predictor of DLB was the presence of soft extrapyramidal signs (mean UPDRS score: 4.04 ± 5.9) detected in 72% of patients, followed by REM sleep behavior disorder (60%) and fluctuations (60%). Wrong performances in the pentagon's number of angles were obtained in 44% of DLB and 3.7% of AD patients and correlated with speed of visual attention. Executive functions, visual attention and visuospatial abilities were worse in DLB, while verbal episodic memory impairment was greater in AD. Deficits in the visual-perceptual domain were present in both MCI-DLB and AD. Conclusions: Poor performance in the pentagon's number of angles is specific of DLB and correlates with speed of visual attention. The dorsal visual stream seems specifically more impaired in MCI-DLB with respect to the ventral visual stream, the latter being involved in both DLB and AD. These cognitive features, associated with subtle extrapyramidal signs, should alert clinicians to a diagnostic hypothesis of DLB.
- Published
- 2015
18. Applying Flow Virometry to Study the HIV Envelope Glycoprotein and Differences Across HIV Model Systems.
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Burnie J, Fernandes C, Patel A, Persaud AT, Chaphekar D, Wei D, Lee TKH, Tang VA, Cicala C, Arthos J, and Guzzo C
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- Humans, HEK293 Cells, HIV Antibodies immunology, Antibodies, Monoclonal immunology, CD4-Positive T-Lymphocytes virology, CD4-Positive T-Lymphocytes immunology, HIV Infections virology, env Gene Products, Human Immunodeficiency Virus chemistry, env Gene Products, Human Immunodeficiency Virus metabolism, env Gene Products, Human Immunodeficiency Virus genetics, env Gene Products, Human Immunodeficiency Virus immunology, HIV-1 genetics, HIV-1 physiology, HIV-1 immunology, Virion metabolism
- Abstract
The HIV envelope glycoprotein (Env) is a trimeric protein that facilitates viral binding and fusion with target cells. As the sole viral protein on the HIV surface, Env is important both for immune responses to HIV and in vaccine designs. Targeting Env in clinical applications is challenging due to its heavy glycosylation, high genetic variability, conformational camouflage, and its low abundance on virions. Thus, there is a critical need to better understand this protein. Flow virometry (FV) is a useful methodology for phenotyping the virion surface in a high-throughput, single virion manner. To demonstrate the utility of FV to characterize Env, we stained HIV virions with a panel of 85 monoclonal antibodies targeting different regions of Env. A broad range of antibodies yielded robust staining of Env, with V3 antibodies showing the highest quantitative staining. A subset of antibodies tested in parallel on viruses produced in CD4
+ T cell lines, HEK293T cells, and primary cells showed that the cellular model of virus production can impact Env detection. Finally, in addition to being able to highlight Env heterogeneity on virions, we show FV can sensitively detect differences in Env conformation when soluble CD4 is added to virions before staining.- Published
- 2024
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19. Virion-incorporated CD14 enables HIV-1 to bind LPS and initiate TLR4 signaling in immune cells.
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Persaud AT, Khela J, Fernandes C, Chaphekar D, Burnie J, Tang VA, Colpitts CC, and Guzzo C
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- Humans, Chemokine CCL5 metabolism, Monocytes metabolism, Monocytes immunology, Monocytes virology, Signal Transduction, THP-1 Cells, Toll-Like Receptor 4 metabolism, Tumor Necrosis Factor-alpha metabolism, HIV Infections virology, HIV Infections immunology, HIV Infections metabolism, HIV-1 immunology, HIV-1 physiology, Lipopolysaccharide Receptors metabolism, Lipopolysaccharides metabolism, Virion metabolism
- Abstract
HIV-1 has a broad range of nuanced interactions with the immune system, and the incorporation of cellular proteins by nascent virions continues to redefine our understanding of the virus-host relationship. Proteins located at the sites of viral egress can be selectively incorporated into the HIV-1 envelope, imparting new functions and phenotypes onto virions, and impacting viral spread and disease. Using virion capture assays and western blot, we show that HIV-1 can incorporate the myeloid antigen CD14 into its viral envelope. Virion-incorporated CD14 remained biologically active and able to bind its natural ligand, bacterial lipopolysaccharide (LPS), as demonstrated by flow virometry and immunoprecipitation assays. Using a Toll-like receptor 4 (TLR4) reporter cell line, we also demonstrated that virions with bound LPS can trigger TLR4 signaling to activate transcription factors that regulate inflammatory gene expression. Complementary assays with THP-1 monocytes demonstrated enhanced secretion of inflammatory cytokines like tumor necrosis factor alpha (TNF-α) and the C-C chemokine ligand 5 (CCL5), when exposed to LPS-loaded virus. These data highlight a new type of interplay between HIV-1 and the myeloid cell compartment, a previously well-established cellular contributor to HIV-1 pathogenesis and inflammation. Persistent gut inflammation is a hallmark of chronic HIV-1 infection, and contributing to this effect is the translocation of microbes across the gut epithelium. Our data herein provide proof of principle that virion-incorporated CD14 could be a novel mechanism through which HIV-1 can drive chronic inflammation, facilitated by HIV-1 particles binding bacterial LPS and initiating inflammatory signaling in TLR4-expressing cells.IMPORTANCEHIV-1 establishes a lifelong infection accompanied by numerous immunological changes. Inflammation of the gut epithelia, exacerbated by the loss of mucosal T cells and cytokine dysregulation, persists during HIV-1 infection. Feeding back into this loop of inflammation is the translocation of intestinal microbes across the gut epithelia, resulting in the systemic dissemination of bacterial antigens, like lipopolysaccharide (LPS). Our group previously demonstrated that the LPS receptor, CD14, can be readily incorporated by HIV-1 particles, supporting previous clinical observations of viruses derived from patient plasma. We now show that CD14 can be incorporated by several primary HIV-1 isolates and that this virion-incorporated CD14 can remain functional, enabling HIV-1 to bind to LPS. This subsequently allowed CD14
+ virions to transfer LPS to monocytic cells, eliciting pro-inflammatory signaling and cytokine secretion. We posit here that virion-incorporated CD14 is a potential contributor to the dysregulated immune responses present in the setting of HIV-1 infection., Competing Interests: The authors declare no conflict of interest.- Published
- 2024
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20. Identification of CD38, CD97, and CD278 on the HIV surface using a novel flow virometry screening assay.
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Burnie J, Fernandes C, Chaphekar D, Wei D, Ahmed S, Persaud AT, Khader N, Cicala C, Arthos J, Tang VA, and Guzzo C
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- Humans, Virion genetics, Cell Line, Viruses, HIV Infections metabolism
- Abstract
While numerous cellular proteins in the HIV envelope are known to alter virus infection, methodology to rapidly phenotype the virion surface in a high throughput, single virion manner is lacking. Thus, many human proteins may exist on the virion surface that remain undescribed. Herein, we developed a novel flow virometry screening assay to discover new proteins on the surface of HIV particles. By screening a CD4
+ T cell line and its progeny virions, along with four HIV isolates produced in primary cells, we discovered 59 new candidate proteins in the HIV envelope that were consistently detected across diverse HIV isolates. Among these discoveries, CD38, CD97, and CD278 were consistently present at high levels on virions when using orthogonal techniques to corroborate flow virometry results. This study yields new discoveries about virus biology and demonstrates the utility and feasibility of a novel flow virometry assay to phenotype individual virions., (© 2023. The Author(s).)- Published
- 2023
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21. SARS-CoV-2 Nucleocapsid Protein is Associated With Lower Testosterone Levels: An Experimental Study.
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Lucio Carrasco CH, Noda P, Barbosa AP, Vieira Borges da Silva EK, Gasque Bomfim C, Ventura Fernandes BH, Teixeira TA, Nunes Duarte Neto A, Nascimento Saldiva PH, Achoa Filho K, Rodrigues Guzzo C, Durigon EL, Affonso Fonseca FL, Corazzini R, Fanelli C, Noronha IL, and Hallak J
- Abstract
The ongoing COVID-19 pandemic represents an extra burden in the majority of public and private health systems worldwide beyond the most pessimistic expectations, driving an urgent rush to develop effective vaccines and effective medical treatments against the SARS-CoV-2 pandemic. The Nucleocapsid structural viral protein is remarkably immunogenic and hugely expressed during infection. High IgG antibodies against Nucleocapsid protein (N protein) levels were detected in the serum of COVID-19 patients, confirming its pivotal antigen role for a T lymphocyte response in a vaccine microenvironment. Currently, adverse events associated with immunizations have raised some degree of concern, irrespective of its huge benefits in dealing with disease severity and decreasing mortality and morbidity. This hitherto study evaluates histological changes in rats' testes, epididymis, prostate, and seminal vesicles and analyzes hormone levels after solely N protein inoculation. Therefore, we exposed a group of Lewis rats to weekly injections of the recombinant N protein for 28 days, while a control group was inoculated with a buffer solution. The N group revealed a more significant number of spermatozoa. Spermatozoa in the seminiferous tubules were counted in twenty 400 × microscopy fields (mean of 9.2 vs. 4.6 in the control group; p < 0,01), but significantly lower testosterone levels (mean of 125.70 ng/dl vs. 309,00 ng/dl in the control group; p < 0,05) were found. No other histological and biochemical changes were displayed. Conclusively, these data suggest testicular hormonal imbalance mediated by the SARS-CoV-2 N protein that could be linked to reported post-COVID-19 syndrome hypogonadism. More relevant research might be performed to confirm this viral antigen's deleterious mechanism in the human testicular microenvironment, particular in Leydig cell function., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Lucio Carrasco, Noda, Barbosa, Vieira Borges da Silva, Gasque Bomfim, Ventura Fernandes, Teixeira, Nunes Duarte Neto, Nascimento Saldiva, Achoa Filho, Rodrigues Guzzo, Durigon, Affonso Fonseca, Corazzini, Fanelli, Noronha and Hallak.)
- Published
- 2022
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22. P-selectin glycoprotein ligand-1 (PSGL-1/CD162) is incorporated into clinical HIV-1 isolates and can mediate virus capture and subsequent transfer to permissive cells.
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Burnie J, Persaud AT, Thaya L, Liu Q, Miao H, Grabinsky S, Norouzi V, Lusso P, Tang VA, and Guzzo C
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- Antiviral Agents metabolism, DNA metabolism, Humans, Leukocytes, Mononuclear, Membrane Glycoproteins, HIV Infections, HIV-1 genetics, HIV-1 metabolism, P-Selectin metabolism
- Abstract
Background: P-selectin glycoprotein ligand-1 (PSGL-1/CD162) has been studied extensively for its role in mediating leukocyte rolling through interactions with its cognate receptor, P-selectin. Recently, PSGL-1 was identified as a novel HIV-1 host restriction factor, particularly when expressed at high levels in the HIV envelope. Importantly, while the potent antiviral activity of PSGL-1 has been clearly demonstrated in various complementary model systems, the breadth of PSGL-1 incorporation across genetically diverse viral isolates and clinical isolates has yet to be described. Additionally, the biological activity of virion-incorporated PSGL-1 has also yet to be shown., Results: Herein we assessed the levels of PSGL-1 on viruses produced through transfection with various amounts of PSGL-1 plasmid DNA (0-250 ng), compared to levels of PSGL-1 on viruses produced through infection of T cell lines and primary PBMC. We found that very low levels of PSGL-1 plasmid DNA (< 2.5 ng/well) were necessary to generate virus models that could closely mirror the phenotype of viruses produced via infection of T cells and PBMC. Unique to this study, we show that PSGL-1 is incorporated in a broad range of HIV-1 and SIV isolates and that virions with incorporated PSGL-1 are detectable in plasma from viremic HIV-1-infected individuals, corroborating the relevance of PSGL-1 in natural infection. Additionally, we show that PSGL-1 on viruses can bind its cognate selectin receptors, P-, E-, and L-selectins. Finally, we show viruses with endogenous levels of PSGL-1 can be captured by P-selectin and transferred to HIV-permissive bystander cells, highlighting a novel role for PSGL-1 in HIV-1 infection. Notably, viruses which contained high levels of PSGL-1 were noninfectious in our hands, in line with previous findings reporting the potent antiviral activity of PSGL-1., Conclusions: Our results indicate that levels of PSGL-1 incorporation into virions can vary widely among model systems tested, and that careful tailoring of plasmid levels is required to recapitulate physiological systems when using pseudovirus models. Taken together, our data suggest that PSGL-1 may play diverse roles in the physiology of HIV-1 infection, particularly due to the functionally active state of PSGL-1 on virion surfaces and the breadth of PSGL-1 incorporation among a wide range of viral isolates., (© 2022. The Author(s).)
- Published
- 2022
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23. Human monocytes store and secrete preformed CCL5, independent of de novo protein synthesis.
- Author
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Persaud AT, Bennett SA, Thaya L, Burnie J, and Guzzo C
- Subjects
- Cytokines metabolism, Humans, Protein Biosynthesis, Chemokine CCL5 metabolism, Leukocytes, Mononuclear metabolism, Monocytes metabolism
- Abstract
Monocytes are a subset of circulating peripheral blood mononuclear cells with diverse roles in immunity, including sentinel roles in cytokine secretion. Conventionally, cytokines require an inductive stimulus for their expression and secretion, resulting in a time lag from the time of stimulation to when the proteins are packaged and secreted. Because cytokines are the main communicators in the immune system, their temporal expression is a key factor in coordinating responses to efficiently resolve infection. Herein, we identify that circulating human monocytes contain preformed cytokines that are stored intracellularly, in both resting and activated states. Having preformed cytokines bypasses the time lag associated with de novo synthesis, allowing monocytes to secrete immune mediators immediately upon activation or sensing of microbe-associated molecular patterns. We demonstrate here that, out of several cytokines evaluated, human monocytes contain a previously undescribed reservoir of the preformed chemokine CCL5. Furthermore, we showed that CCL5 could be secreted from monocytes treated with the protein synthesis inhibitor (cycloheximide) and Golgi blocker (brefeldin A). We examined the possibility for uptake of extracellular CCL5 from platelet aggregates and observed no significant levels of platelet binding to our enriched monocyte preparations, indicating that the source of preformed CCL5 was not from platelets. Preformed CCL5 was observed to be distributed throughout the cytoplasm and partially colocalized with CD63+ and Rab11A+ membranes, implicating endosomal compartments in the intracellular storage and trafficking of CCL5., (©2021 Society for Leukocyte Biology.)
- Published
- 2022
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24. A UV-LED module that is highly effective at inactivating human coronaviruses and HIV-1.
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Persaud AT, Burnie J, Thaya L, DSouza L, Martin S, and Guzzo C
- Subjects
- Humans, Coronavirus 229E, Human radiation effects, Disinfection methods, HIV-1 radiation effects, Ultraviolet Rays, Virus Inactivation radiation effects
- Abstract
Ultraviolet (UV) light has previously been established as useful method of disinfection, with demonstrated efficacy to inactivate a broad range of microorganisms. The advent of ultraviolet light-emitting diodes provides advantages in ease of disinfection, in that there can be delivery of germicidal UV with the same light unit that delivers standard white light to illuminate a room. Herein we demonstrate the efficacy and feasibility of ultraviolet light-emitting diodes as a means of decontamination by inactivating two distinct virus models, human coronavirus 229E and human immunodeficiency virus. Importantly, the same dose of ultraviolet light that inactivated human viruses also elicited complete inactivation of ultraviolet-resistant bacterial spores (Bacillus pumilus), a gold standard for demonstrating ultraviolet-mediated disinfection. This work demonstrates that seconds of ultraviolet light-emitting diodes (UV-LED) exposure can inactivate viruses and bacteria, highlighting that UV-LED could be a useful and practical tool for broad sanitization of public spaces., (© 2022. The Author(s).)
- Published
- 2022
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25. Engineering pan-HIV-1 neutralization potency through multispecific antibody avidity.
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Rujas E, Cui H, Burnie J, Aschner CB, Zhao T, Insausti S, Muthuraman K, Semesi A, Ophel J, Nieva JL, Seaman MS, Guzzo C, Treanor B, and Julien JP
- Subjects
- Antibodies, Neutralizing chemistry, Broadly Neutralizing Antibodies chemistry, Broadly Neutralizing Antibodies immunology, Epitopes chemistry, Epitopes immunology, HIV Antibodies chemistry, HIV Antibodies genetics, HIV-1 immunology, Humans, Models, Molecular, Protein Conformation, Structure-Activity Relationship, Antibodies, Neutralizing immunology, Antibody Affinity immunology, HIV Antibodies immunology, Neutralization Tests methods, Protein Engineering methods
- Abstract
Deep mining of B cell repertoires of HIV-1-infected individuals has resulted in the isolation of dozens of HIV-1 broadly neutralizing antibodies (bNAbs). Yet, it remains uncertain whether any such bNAbs alone are sufficiently broad and potent to deploy therapeutically. Here, we engineered HIV-1 bNAbs for their combination on a single multispecific and avid molecule via direct genetic fusion of their Fab fragments to the human apoferritin light chain. The resulting molecule demonstrated a remarkable median IC
50 value of 0.0009 µg/mL and 100% neutralization coverage of a broad HIV-1 pseudovirus panel (118 isolates) at a 4 µg/mL cutoff-a 32-fold enhancement in viral neutralization potency compared to a mixture of the corresponding HIV-1 bNAbs. Importantly, Fc incorporation on the molecule and engineering to modulate Fc receptor binding resulted in IgG-like bioavailability in vivo. This robust plug-and-play antibody design is relevant against indications where multispecificity and avidity are leveraged simultaneously to mediate optimal biological activity., Competing Interests: Competing interest statement: The Hospital for Sick Children has applied for patents concerning the Multabody platform technology that are related to this work. B.T. and J.-P.J. are founders of Radiant Biotherapeutics and are members of its Scientific Advisory Board., (Copyright © 2022 the Author(s). Published by PNAS.)- Published
- 2022
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26. Functional Anatomy of the Trimer Apex Reveals Key Hydrophobic Constraints That Maintain the HIV-1 Envelope Spike in a Closed State.
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Zhang P, Kwon AL, Guzzo C, Liu Q, Schmeisser H, Miao H, Lin Y, Cimbro R, Huang J, Connors M, Schmidt SD, Dolan MA, Armstrong AA, and Lusso P
- Subjects
- Amino Acid Motifs, Amino Acid Sequence, Antibodies, Neutralizing immunology, HIV Antibodies, HIV Infections virology, HIV-1 chemistry, HIV-1 immunology, Humans, Hydrophobic and Hydrophilic Interactions, Mutation, env Gene Products, Human Immunodeficiency Virus genetics, HIV-1 genetics, env Gene Products, Human Immunodeficiency Virus chemistry, env Gene Products, Human Immunodeficiency Virus immunology
- Abstract
The human immunodeficiency virus type 1 (HIV-1) envelope trimer maintains a closed, metastable configuration to protect vulnerable epitopes from neutralizing antibodies. Here, we identify key hydrophobic constraints at the trimer apex that function as global stabilizers of the HIV-1 envelope spike configuration. Mutation of individual residues within four hydrophobic clusters that fasten together the V1V2, V3, and C4 domains at the apex of gp120 dramatically increases HIV-1 sensitivity to weak and restricted neutralizing antibodies targeting epitopes that are largely concealed in the prefusion Env spike, consistent with the adoption of a partially open trimer configuration. Conversely, the same mutations decrease the sensitivity to broad and potent neutralizing antibodies that preferentially recognize the closed trimer. Sera from chronically HIV-infected patients neutralize open mutants with enhanced potency, compared to the wild-type virus, suggesting that a large fraction of host-generated antibodies target concealed epitopes. The identification of structural constraints that maintain the HIV-1 envelope in an antibody-protected state may inform the design of a protective vaccine. IMPORTANCE Elucidating the structure and function of the HIV-1 envelope proteins is critical for the design of an effective vaccine. Despite the availability of many high-resolution structures, key functional correlates in the envelope trimer remain undefined. We utilized a combination of structural analysis, in silico energy calculation, mutagenesis, and neutralization profiling to dissect the functional anatomy of the trimer apex, which acts as a global regulator of the HIV-1 spike conformation. We identify four hydrophobic clusters that stabilize the spike in a tightly closed configuration and, thereby, play a critical role in protecting it from the reach of neutralizing antibodies., (Copyright © 2021 Zhang et al.)
- Published
- 2021
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27. Flow Virometry Quantification of Host Proteins on the Surface of HIV-1 Pseudovirus Particles.
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Burnie J, Tang VA, Welsh JA, Persaud AT, Thaya L, Jones JC, and Guzzo C
- Subjects
- Biomarkers, HIV-1 genetics, Humans, Reproducibility of Results, Flow Cytometry methods, Genetic Engineering, HIV-1 metabolism, Host-Pathogen Interactions, Viral Load methods, Virion genetics, Virion metabolism
- Abstract
The HIV-1 glycoprotein spike (gp120) is typically the first viral antigen that cells encounter before initiating immune responses, and is often the sole target in vaccine designs. Thus, characterizing the presence of cellular antigens on the surfaces of HIV particles may help identify new antiviral targets or impact targeting of gp120. Despite the importance of characterizing proteins on the virion surface, current techniques available for this purpose do not support high-throughput analysis of viruses, and typically only offer a semi-quantitative assessment of virus-associated proteins. Traditional bulk techniques often assess averages of viral preparations, which may mask subtle but important differences in viral subsets. On the other hand, microscopy techniques, which provide detail on individual virions, are difficult to use in a high-throughput manner and have low levels of sensitivity for antigen detection. Flow cytometry is a technique that traditionally has been used for rapid, high-sensitivity characterization of single cells, with limited use in detecting viruses, since the small size of viral particles hinders their detection. Herein, we report the detection and surface antigen characterization of HIV-1 pseudovirus particles by light scattering and fluorescence with flow cytometry, termed flow virometry for its specific application to viruses. We quantified three cellular proteins (integrin α4β7, CD14, and CD162/PSGL-1) in the viral envelope by directly staining virion-containing cell supernatants without the requirement of additional processing steps to distinguish virus particles or specific virus purification techniques. We also show that two antigens can be simultaneously detected on the surface of individual HIV virions, probing for the tetraspanin marker, CD81, in addition to α4β7, CD14, and CD162/PSGL-1. This study demonstrates new advances in calibrated flow virometry as a tool to provide sensitive, high-throughput characterization of the viral envelope in a more efficient, quantitative manner than previously reported techniques.
- Published
- 2020
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28. Single-cell sequencing reveals clonal expansions of pro-inflammatory synovial CD8 T cells expressing tissue-homing receptors in psoriatic arthritis.
- Author
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Penkava F, Velasco-Herrera MDC, Young MD, Yager N, Nwosu LN, Pratt AG, Lara AL, Guzzo C, Maroof A, Mamanova L, Cole S, Efremova M, Simone D, Filer A, Brown CC, Croxford AL, Isaacs JD, Teichmann S, Bowness P, Behjati S, and Hussein Al-Mossawi M
- Subjects
- Arthritis, Psoriatic blood, CD8-Positive T-Lymphocytes metabolism, Gene Expression Profiling, Humans, Immunologic Memory, Receptors, Antigen, T-Cell, alpha-beta genetics, Receptors, Antigen, T-Cell, alpha-beta metabolism, Receptors, Chemokine metabolism, Receptors, Lymphocyte Homing genetics, Single-Cell Analysis, Synovial Membrane immunology, Arthritis, Psoriatic immunology, CD8-Positive T-Lymphocytes immunology, Clonal Selection, Antigen-Mediated, Receptors, Lymphocyte Homing metabolism, Synovial Fluid immunology
- Abstract
Psoriatic arthritis (PsA) is a debilitating immune-mediated inflammatory arthritis of unknown pathogenesis commonly affecting patients with skin psoriasis. Here we use complementary single-cell approaches to study leukocytes from PsA joints. Mass cytometry demonstrates a 3-fold expansion of memory CD8 T cells in the joints of PsA patients compared to peripheral blood. Meanwhile, droplet-based and plate-based single-cell RNA sequencing of paired T cell receptor alpha and beta chain sequences show pronounced CD8 T cell clonal expansions within the joints. Transcriptome analyses find these expanded synovial CD8 T cells to express cycling, activation, tissue-homing and tissue residency markers. T cell receptor sequence comparison between patients identifies clonal convergence. Finally, chemokine receptor CXCR3 is upregulated in the expanded synovial CD8 T cells, while two CXCR3 ligands, CXCL9 and CXCL10, are elevated in PsA synovial fluid. Our data thus provide a quantitative molecular insight into the cellular immune landscape of psoriatic arthritis.
- Published
- 2020
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29. The Incorporation of Host Proteins into the External HIV-1 Envelope.
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Burnie J and Guzzo C
- Subjects
- CD4-Positive T-Lymphocytes virology, Humans, Integrins genetics, Macrophages virology, Membrane Proteins genetics, Virion genetics, Virus Release, env Gene Products, Human Immunodeficiency Virus genetics, HIV-1 genetics, Host Microbial Interactions, Membrane Proteins chemistry, env Gene Products, Human Immunodeficiency Virus chemistry
- Abstract
The incorporation of biologically active host proteins into HIV-1 is a well-established phenomenon, particularly due to the budding mechanism of viral egress in which viruses acquire their external lipid membrane directly from the host cell. While this mechanism might seemingly imply that host protein incorporation is a passive uptake of all cellular antigens associated with the plasma membrane at the site of budding, this is not the case. Herein, we review the evidence indicating that host protein incorporation can be a selective and conserved process. We discuss how HIV-1 virions displaying host proteins on their surface can exhibit a myriad of altered phenotypes, with notable impacts on infectivity, homing, neutralization, and pathogenesis. This review describes the canonical and emerging methods to detect host protein incorporation, highlights the well-established host proteins that have been identified on HIV-1 virions, and reflects on the role of these incorporated proteins in viral pathogenesis and therapeutic targeting. Despite many advances in HIV treatment and prevention, there remains a global effort to develop increasingly effective anti-HIV therapies. Given the broad range of biologically active host proteins acquired on the surface of HIV-1, additional studies on the mechanisms and impacts of these incorporated host proteins may inform the development of novel treatments and vaccine designs.
- Published
- 2019
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30. Structural Constraints at the Trimer Apex Stabilize the HIV-1 Envelope in a Closed, Antibody-Protected Conformation.
- Author
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Guzzo C, Zhang P, Liu Q, Kwon AL, Uddin F, Wells AI, Schmeisser H, Cimbro R, Huang J, Doria-Rose N, Schmidt SD, Dolan MA, Connors M, Mascola JR, and Lusso P
- Subjects
- DNA Mutational Analysis, HIV Envelope Protein gp120 chemistry, HIV Envelope Protein gp120 genetics, HIV Infections immunology, HIV-1 genetics, Humans, Immune Evasion, Neutralization Tests, Protein Structure, Quaternary, Antibodies, Neutralizing immunology, HIV Antibodies immunology, HIV Envelope Protein gp120 immunology, HIV-1 immunology
- Abstract
The human immunodeficiency virus type 1 (HIV-1) envelope (Env) trimer evades antibody recognition by adopting a closed prefusion conformation. Here, we show that two conserved tyrosines (Y173, Y177) within the second variable (V2) loop of the gp120 Env glycoprotein are key regulators of the closed, antibody-protected state of the trimer by establishing intramolecular interaction with the base of the third variable (V3) loop. Mutation of Y177 and/or Y173 to phenylalanine or alanine dramatically altered the susceptibility of diverse HIV-1 strains to neutralization, increasing sensitivity to weakly and nonneutralizing antibodies directed against diverse Env regions, consistent with the adoption of an open trimer configuration. Conversely, potent broadly neutralizing antibodies (bNAbs) against different supersites of HIV-1 vulnerability exhibited reduced potency against V2 loop tyrosine mutants, consistent with their preferential targeting of the closed trimer. Mutation of V3 loop residues predicted to interact with the V2 loop tyrosines yielded a similar neutralization phenotype. Sera from chronically HIV-1-infected patients contained very high titers of antibodies capable of neutralizing V2 loop tyrosine mutants but not wild-type viruses, indicating that the bulk of antibodies produced in infected hosts are unable to penetrate the protective shield of the closed trimer. These results identify the tyrosine-mediated V2-V3 loop complex at the trimer apex as a key structural constraint that facilitates HIV-1 evasion from the bulk of host antibodies. IMPORTANCE The extraordinary ability of human immunodeficiency virus type 1 (HIV-1) to evade host immunity represents a major obstacle to the development of a protective vaccine. Thus, elucidating the mechanisms whereby HIV-1 protects its external envelope (Env), which is the sole target of virus-neutralizing antibodies, is an essential step toward vaccine design. We identified a key structural element that maintains the HIV-1 Env trimer in a closed, antibody-resistant conformation. A major role is played by two conserved tyrosines at the apex of the Env spike, whose mutation causes a global opening of the trimer structure, exposing multiple concealed targets for neutralizing antibodies. We also found that HIV-infected individuals produce very large amounts of antibodies that neutralize the open Env form; however, the bulk of these antibodies are unable to penetrate the tight defensive shield of the native virus. This work may help to devise new strategies to overcome the viral defensive mechanisms and facilitate the development of an effective HIV-1 vaccine.
- Published
- 2018
- Full Text
- View/download PDF
31. Recurrent intragenic rearrangements of EGFR and BRAF in soft tissue tumors of infants.
- Author
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Wegert J, Vokuhl C, Collord G, Del Castillo Velasco-Herrera M, Farndon SJ, Guzzo C, Jorgensen M, Anderson J, Slater O, Duncan C, Bausenwein S, Streitenberger H, Ziegler B, Furtwängler R, Graf N, Stratton MR, Campbell PJ, Jones DT, Koelsche C, Pfister SM, Mifsud W, Sebire N, Sparber-Sauer M, Koscielniak E, Rosenwald A, Gessler M, and Behjati S
- Subjects
- Female, Gene Rearrangement, Humans, Infant, Infant, Newborn, Male, Fibrosarcoma genetics, Genes, erbB-1, Kidney Neoplasms genetics, Nephroma, Mesoblastic genetics, Proto-Oncogene Proteins B-raf genetics
- Abstract
Soft tissue tumors of infancy encompass an overlapping spectrum of diseases that pose unique diagnostic and clinical challenges. We studied genomes and transcriptomes of cryptogenic congenital mesoblastic nephroma (CMN), and extended our findings to five anatomically or histologically related soft tissue tumors: infantile fibrosarcoma (IFS), nephroblastomatosis, Wilms tumor, malignant rhabdoid tumor, and clear cell sarcoma of the kidney. A key finding is recurrent mutation of EGFR in CMN by internal tandem duplication of the kinase domain, thus delineating CMN from other childhood renal tumors. Furthermore, we identify BRAF intragenic rearrangements in CMN and IFS. Collectively these findings reveal novel diagnostic markers and therapeutic strategies and highlight a prominent role of isolated intragenic rearrangements as drivers of infant tumors.
- Published
- 2018
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32. Interdomain Stabilization Impairs CD4 Binding and Improves Immunogenicity of the HIV-1 Envelope Trimer.
- Author
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Zhang P, Gorman J, Geng H, Liu Q, Lin Y, Tsybovsky Y, Go EP, Dey B, Andine T, Kwon A, Patel M, Gururani D, Uddin F, Guzzo C, Cimbro R, Miao H, McKee K, Chuang GY, Martin L, Sironi F, Malnati MS, Desaire H, Berger EA, Mascola JR, Dolan MA, Kwong PD, and Lusso P
- Subjects
- AIDS Vaccines immunology, Animals, Antibodies, Neutralizing immunology, Female, HEK293 Cells, HIV Antibodies immunology, HIV Antigens chemistry, HIV Antigens immunology, HIV Envelope Protein gp120 chemistry, HIV Envelope Protein gp120 genetics, HIV Envelope Protein gp120 immunology, HIV Envelope Protein gp120 metabolism, HIV Envelope Protein gp160 chemistry, HIV Envelope Protein gp160 immunology, HIV Envelope Protein gp160 metabolism, HIV-1 genetics, HIV-1 pathogenicity, Humans, Immunization, Models, Molecular, Protein Conformation, Rabbits, Virus Internalization, env Gene Products, Human Immunodeficiency Virus genetics, CD4 Antigens immunology, CD4 Antigens metabolism, HIV-1 immunology, Protein Binding immunology, Protein Domains immunology, Protein Stability, env Gene Products, Human Immunodeficiency Virus chemistry, env Gene Products, Human Immunodeficiency Virus immunology
- Abstract
The HIV-1 envelope (Env) spike is a trimer of gp120/gp41 heterodimers that mediates viral entry. Binding to CD4 on the host cell membrane is the first essential step for infection but disrupts the native antigenic state of Env, posing a key obstacle to vaccine development. We locked the HIV-1 Env trimer in a pre-fusion configuration, resulting in impaired CD4 binding and enhanced binding to broadly neutralizing antibodies. This design was achieved via structure-guided introduction of neo-disulfide bonds bridging the gp120 inner and outer domains and was successfully applied to soluble trimers and native gp160 from different HIV-1 clades. Crystallization illustrated the structural basis for CD4-binding impairment. Immunization of rabbits with locked trimers from two different clades elicited neutralizing antibodies against tier-2 viruses with a repaired glycan shield regardless of treatment with a functional CD4 mimic. Thus, interdomain stabilization provides a widely applicable template for the design of Env-based HIV-1 vaccines., (Published by Elsevier Inc.)
- Published
- 2018
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33. Global Identification of Small Ubiquitin-related Modifier (SUMO) Substrates Reveals Crosstalk between SUMOylation and Phosphorylation Promotes Cell Migration.
- Author
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Uzoma I, Hu J, Cox E, Xia S, Zhou J, Rho HS, Guzzo C, Paul C, Ajala O, Goodwin CR, Jeong J, Moore C, Zhang H, Meluh P, Blackshaw S, Matunis M, Qian J, and Zhu H
- Subjects
- Amino Acid Sequence, HeLa Cells, Humans, Phosphorylation, Phosphotyrosine metabolism, Protein Kinases chemistry, Protein Kinases metabolism, Proteomics, Reproducibility of Results, Signal Transduction, Substrate Specificity, Ubiquitin-Protein Ligases metabolism, Cell Movement, Small Ubiquitin-Related Modifier Proteins metabolism, Sumoylation
- Abstract
Proteomics studies have revealed that SUMOylation is a widely used post-translational modification (PTM) in eukaryotes. However, how SUMO E1/2/3 complexes use different SUMO isoforms and recognize substrates remains largely unknown. Using a human proteome microarray-based activity screen, we identified over 2500 proteins that undergo SUMO E3-dependent SUMOylation. We next constructed a SUMO isoform- and E3 ligase-dependent enzyme-substrate relationship network. Protein kinases were significantly enriched among SUMOylation substrates, suggesting crosstalk between phosphorylation and SUMOylation. Cell-based analyses of tyrosine kinase, PYK2, revealed that SUMOylation at four lysine residues promoted PYK2 autophosphorylation at tyrosine 402, which in turn enhanced its interaction with SRC and full activation of the SRC-PYK2 complex. SUMOylation on WT but not the 4KR mutant of PYK2 further elevated phosphorylation of the downstream components in the focal adhesion pathway, such as paxillin and Erk1/2, leading to significantly enhanced cell migration during wound healing. These studies illustrate how our SUMO E3 ligase-substrate network can be used to explore crosstalk between SUMOylation and other PTMs in many biological processes., (© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2018
- Full Text
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34. The driver landscape of sporadic chordoma.
- Author
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Tarpey PS, Behjati S, Young MD, Martincorena I, Alexandrov LB, Farndon SJ, Guzzo C, Hardy C, Latimer C, Butler AP, Teague JW, Shlien A, Futreal PA, Shah S, Bashashati A, Jamshidi F, Nielsen TO, Huntsman D, Baumhoer D, Brandner S, Wunder J, Dickson B, Cogswell P, Sommer J, Phillips JJ, Amary MF, Tirabosco R, Pillay N, Yip S, Stratton MR, Flanagan AM, and Campbell PJ
- Subjects
- Case-Control Studies, Cell Line, Tumor, Class I Phosphatidylinositol 3-Kinases genetics, Class Ia Phosphatidylinositol 3-Kinase, Gene Duplication, Humans, Phosphatidylinositol 3-Kinases genetics, Phosphatidylinositol 3-Kinases metabolism, Polymorphism, Single Nucleotide, Bone Neoplasms genetics, Chordoma genetics, Fetal Proteins genetics, Mutation, T-Box Domain Proteins genetics, Vesicular Transport Proteins genetics
- Abstract
Chordoma is a malignant, often incurable bone tumour showing notochordal differentiation. Here, we defined the somatic driver landscape of 104 cases of sporadic chordoma. We reveal somatic duplications of the notochordal transcription factor brachyury (T) in up to 27% of cases. These variants recapitulate the rearrangement architecture of the pathogenic germline duplications of T that underlie familial chordoma. In addition, we find potentially clinically actionable PI3K signalling mutations in 16% of cases. Intriguingly, one of the most frequently altered genes, mutated exclusively by inactivating mutation, was LYST (10%), which may represent a novel cancer gene in chordoma.Chordoma is a rare often incurable malignant bone tumour. Here, the authors investigate driver mutations of sporadic chordoma in 104 cases, revealing duplications in notochordal transcription factor brachyury (T), PI3K signalling mutations, and mutations in LYST, a potential novel cancer gene in chordoma.
- Published
- 2017
- Full Text
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35. IL-27 enhances LPS-induced IL-1β in human monocytes and murine macrophages.
- Author
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Petes C, Wynick C, Guzzo C, Mehta D, Logan S, Banfield BW, Basta S, Cooper A, and Gee K
- Subjects
- Adenosine Triphosphate pharmacology, Animals, Caspase 1 genetics, Caspase 1 immunology, Cell Line, Tumor, Humans, Interleukin-1beta genetics, Interleukins genetics, Mice, Mice, Knockout, Receptors, Purinergic P2X7 genetics, Receptors, Purinergic P2X7 immunology, Signal Transduction genetics, Signal Transduction immunology, Toll-Like Receptor 4 agonists, Toll-Like Receptor 4 genetics, Toll-Like Receptor 4 immunology, Interleukin-1beta immunology, Interleukins immunology, Lipopolysaccharides pharmacology, Macrophages immunology, Monocytes immunology, Signal Transduction drug effects
- Abstract
IL-27 bridges innate and adaptive immunity by modulating cytokine production from myeloid cells and regulating Th cell differentiation. During bacterial infection, TLR4 triggering by LPS induces IL-27 production by monocytes and macrophages. We have previously shown that IL-27 can prime monocytes for LPS responsiveness by enhancing TLR4 expression and intracellular signaling. If unregulated, this could result in damaging inflammation, whereas on the other hand, this may also provide greater responses by inflammatory processes induced in response to bacterial pathogens. A key process in fine-tuning inflammatory responses is activation of the inflammasome, which ultimately results in IL-1β production. Herein, we investigated the molecular mechanisms by which IL-27 modulates LPS-induced IL-1β secretion in monocytes and macrophages. We found that when delivered simultaneously with LPS, IL-27 augments activation of caspase-1 and subsequent release of IL-1β. Furthermore, we determined that IL-27 primes cells for enhanced IL-1β production by up-regulating surface expression of TLR4 and P2X purinoceptor 7 (P2X7) for enhanced LPS and ATP signaling, respectively. These findings provide new evidence that IL-27 plays an important role in the proinflammatory capacity of monocytes and macrophages via enhancing IL-1β secretion levels triggered by dual LPS-ATP stimulation., (© Society for Leukocyte Biology.)
- Published
- 2017
- Full Text
- View/download PDF
36. Corrigendum: Quaternary contact in the initial interaction of CD4 with the HIV-1 envelope trimer.
- Author
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Liu Q, Acharya P, Dolan MA, Zhang P, Guzzo C, Lu J, Kwon A, Gururani D, Miao H, Bylund T, Chuang GY, Druz A, Zhou T, Rice WJ, Wigge C, Carragher B, Potter CS, Kwong PD, and Lusso P
- Published
- 2017
- Full Text
- View/download PDF
37. Quaternary contact in the initial interaction of CD4 with the HIV-1 envelope trimer.
- Author
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Liu Q, Acharya P, Dolan MA, Zhang P, Guzzo C, Lu J, Kwon A, Gururani D, Miao H, Bylund T, Chuang GY, Druz A, Zhou T, Rice WJ, Wigge C, Carragher B, Potter CS, Kwong PD, and Lusso P
- Subjects
- Amino Acid Sequence, Antibodies, Neutralizing chemistry, Antibodies, Neutralizing metabolism, Binding Sites, CD4 Antigens ultrastructure, Cryoelectron Microscopy, HEK293 Cells, HIV Antibodies chemistry, HIV Antibodies metabolism, HIV Envelope Protein gp120 ultrastructure, HIV Infections metabolism, Humans, Kinetics, Mutagenesis, Protein Binding, Protein Stability, Protein Structure, Quaternary, Surface Plasmon Resonance, CD4 Antigens chemistry, CD4 Antigens metabolism, HIV Envelope Protein gp120 chemistry, HIV Envelope Protein gp120 metabolism, HIV-1 metabolism, Protein Multimerization
- Abstract
Binding of the gp120 envelope (Env) glycoprotein to the CD4 receptor is the first step in the HIV-1 infectious cycle. Although the CD4-binding site has been extensively characterized, the initial receptor interaction has been difficult to study because of major CD4-induced structural rearrangements. Here we used cryogenic electron microscopy (cryo-EM) to visualize the initial contact of CD4 with the HIV-1 Env trimer at 6.8-Å resolution. A single CD4 molecule is embraced by a quaternary HIV-1-Env surface formed by coalescence of the previously defined CD4-contact region with a second CD4-binding site (CD4-BS2) in the inner domain of a neighboring gp120 protomer. Disruption of CD4-BS2 destabilized CD4-trimer interaction and abrogated HIV-1 infectivity by preventing the acquisition of coreceptor-binding competence. A corresponding reduction in HIV-1 infectivity occurred after the mutation of CD4 residues that interact with CD4-BS2. Our results document the critical role of quaternary interactions in the initial HIV-Env-receptor contact, with implications for treatment and vaccine design.
- Published
- 2017
- Full Text
- View/download PDF
38. Tyrosine-sulfated V2 peptides inhibit HIV-1 infection via coreceptor mimicry.
- Author
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Cimbro R, Peterson FC, Liu Q, Guzzo C, Zhang P, Miao H, Van Ryk D, Ambroggio X, Hurt DE, De Gioia L, Volkman BF, Dolan MA, and Lusso P
- Subjects
- Amino Acid Sequence, Anti-HIV Agents chemistry, Anti-HIV Agents metabolism, Anti-HIV Agents pharmacology, Binding Sites, CD4 Antigens chemistry, CD4 Antigens metabolism, HIV Envelope Protein gp120 chemistry, HIV Infections drug therapy, HIV-1 drug effects, Humans, Molecular Docking Simulation, Molecular Dynamics Simulation, Nuclear Magnetic Resonance, Biomolecular, Peptide Fragments chemistry, Peptide Fragments pharmacology, Protein Binding, Protein Conformation, Receptors, CCR5 chemistry, Tyrosine analogs & derivatives, Tyrosine chemistry, HIV Envelope Protein gp120 metabolism, HIV Infections metabolism, HIV Infections virology, HIV-1 physiology, Molecular Mimicry, Peptide Fragments metabolism, Receptors, CCR5 metabolism
- Abstract
Tyrosine sulfation is a post-translational modification that facilitates protein-protein interaction. Two sulfated tyrosines (Tys173 and Tys177) were recently identified within the second variable (V2) loop of the major HIV-1 envelope glycoprotein, gp120, and shown to contribute to stabilizing the intramolecular interaction between V2 and the third variable (V3) loop. Here, we report that tyrosine-sulfated peptides derived from V2 act as structural and functional mimics of the CCR5 N-terminus and potently block HIV-1 infection. Nuclear magnetic and surface plasmon resonance analyses indicate that a tyrosine-sulfated V2 peptide (pV2α-Tys) adopts a CCR5-like helical conformation and directly interacts with gp120 in a CD4-dependent fashion, competing with a CCR5 N-terminal peptide. Sulfated V2 mimics, but not their non-sulfated counterparts, inhibit HIV-1 entry and fusion by preventing coreceptor utilization, with the highly conserved C-terminal sulfotyrosine, Tys177, playing a dominant role. Unlike CCR5 N-terminal peptides, V2 mimics inhibit a broad range of HIV-1 strains irrespective of their coreceptor tropism, highlighting the overall structural conservation of the coreceptor-binding site in gp120. These results document the use of receptor mimicry by a retrovirus to occlude a key neutralization target site and provide leads for the design of therapeutic strategies against HIV-1., (Published by Elsevier B.V.)
- Published
- 2016
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39. Clinical and Cognitive Phenotype of Mild Cognitive Impairment Evolving to Dementia with Lewy Bodies.
- Author
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Cagnin A, Bussè C, Gardini S, Jelcic N, Guzzo C, Gnoato F, Mitolo M, Ermani M, and Caffarra P
- Abstract
Objective: The aim of this study was to determine which characteristics could better distinguish dementia with Lewy bodies (DLB) from Alzheimer's disease (AD) at the mild cognitive impairment (MCI) stage, with particular emphasis on visual space and object perception abilities., Methods: Fifty-three patients with mild cognitive deficits that were eventually diagnosed with probable DLB (MCI-DLB: n = 25) and AD (MCI-AD: n = 28) at a 3-year follow-up were retrospectively studied. At the first visit, the patients underwent cognitive assessment including the Qualitative Scoring Mini Mental State Examination Pentagon Test and the Visual Object and Space Perception Battery. The Neuropsychiatric Inventory Questionnaire, Unified Parkinson's Disease Rating Scale (UPDRS) and questionnaires for cognitive fluctuations and sleep disorders were also administered., Results: The best clinical predictor of DLB was the presence of soft extrapyramidal signs (mean UPDRS score: 4.04 ± 5.9) detected in 72% of patients, followed by REM sleep behavior disorder (60%) and fluctuations (60%). Wrong performances in the pentagon's number of angles were obtained in 44% of DLB and 3.7% of AD patients and correlated with speed of visual attention. Executive functions, visual attention and visuospatial abilities were worse in DLB, while verbal episodic memory impairment was greater in AD. Deficits in the visual-perceptual domain were present in both MCI-DLB and AD., Conclusions: Poor performance in the pentagon's number of angles is specific of DLB and correlates with speed of visual attention. The dorsal visual stream seems specifically more impaired in MCI-DLB with respect to the ventral visual stream, the latter being involved in both DLB and AD. These cognitive features, associated with subtle extrapyramidal signs, should alert clinicians to a diagnostic hypothesis of DLB.
- Published
- 2015
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40. Structural Determinants for the Selective Anti-HIV-1 Activity of the All-β Alternative Conformer of XCL1.
- Author
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Guzzo C, Fox JC, Miao H, Volkman BF, and Lusso P
- Subjects
- Amino Acid Substitution genetics, CD8-Positive T-Lymphocytes immunology, Cells, Cultured, Chemokines, C genetics, Glycosaminoglycans metabolism, HIV-1 growth & development, HIV-1 immunology, Humans, Protein Binding genetics, Protein Folding, Structure-Activity Relationship, Chemokines, C metabolism, HIV Envelope Protein gp120 metabolism, HIV Infections prevention & control, Receptors, G-Protein-Coupled metabolism, Virus Internalization
- Abstract
Unlabelled: HIV-1 replication is regulated in vivo by a complex network of cytokines and chemokines. XCL1/lymphotactin, a unique metamorphic chemokine, was recently identified as a broad-spectrum endogenous HIV-1 inhibitor that blocks viral entry via direct interaction with the gp120 envelope glycoprotein. HIV-1 inhibition by XCL1 requires access to the alternative all-β conformation, which interacts with glycosaminoglycans (GAGs) but not with the specific XCL1 receptor, XCR1. To investigate the structural determinants of the HIV-inhibitory function of XCL1, we performed a detailed structure-function analysis of a stabilized all-β variant, XCL1 W55D. Individual alanine substitutions of two basic residues within the 40s' loop, K42 and R43, abrogated the ability of XCL1 to bind to the viral envelope and block HIV-1 infection; moreover, a loss of HIV-inhibitory function, albeit less marked, was seen upon individual mutation of three additional basic residues: R18, R35, and K46. In contrast, mutation of K42 to arginine did not cause any loss of function, suggesting that the interaction with gp120 is primarily electrostatic in nature. Strikingly, four of these five residues cluster to form a large (∼350 Å(2)) positively charged surface in the all-β XCL1 conformation, whereas they are dissociated in the classic chemokine fold, which is inactive against HIV-1, providing a structural basis for the selective antiviral activity of the alternatively folded XCL1. Furthermore, we observed that changes to the N-terminal domain, which is proximal to the cluster of putative HIV-1 gp120-interacting residues, also affect the antiviral activity of XCL1. Interestingly, the complement of residues involved in HIV-1 blockade is partially overlapping, but distinct from those involved in the GAG-binding function of XCL1. These data identify key structural determinants of anti-HIV activity in XCL1, providing new templates for the development of HIV-1 entry inhibitors., Importance: The host immune system controls HIV-1 infection through a wide array of inhibitory responses, including the induction of cytotoxic effector cells and the secretion of noncytolytic soluble antiviral factors such as cytokines and chemokines. We recently identified XCL1/lymphotactin, a chemokine primarily produced by CD8(+) T cells, as a novel endogenous factor with broad anti-HIV activity. Strikingly, only one of the two conformations that XCL1 can adopt in solution, the alternative all-β fold, mediates antiviral activity. At variance with the classic HIV-inhibitory chemokines such as CCL5/RANTES, XCL1 acts via direct interaction with the external viral envelope glycoprotein, gp120. Here, we identify the interactive surface of XCL1 that is implicated in binding to the HIV-1 envelope and HIV-1 inhibition, providing a structural basis to explain why only the all-β XCL1 conformer is effective against HIV-1. Our findings may be useful in guiding the rational design of new inhibitors of HIV-1 entry., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)
- Published
- 2015
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41. Characterization of HIV-1 entry inhibitors with broad activity against R5 and X4 viral strains.
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Sironi F, Malnati M, Mongelli N, Cozzi P, Guzzo C, Ghezzi S, Martínez-Romero C, García-Sastre A, Lusso P, Jabes D, and Biswas P
- Subjects
- 3T3 Cells, Animals, Antiviral Agents pharmacology, Benzylamines, Cell Death drug effects, Cell Line, Cyclams, Cyclohexanes pharmacology, Flow Cytometry, HIV Envelope Protein gp120 metabolism, HIV Infections pathology, HIV Infections virology, HIV-1 drug effects, Heterocyclic Compounds pharmacology, Humans, Maraviroc, Membrane Fusion drug effects, Mice, Receptors, CCR5 metabolism, Receptors, CXCR4 metabolism, Triazoles pharmacology, Virus Replication drug effects, HIV Fusion Inhibitors pharmacology, HIV-1 physiology, Virus Internalization drug effects
- Abstract
Background: Combined antiretroviral therapy has drastically reduced mortality and morbidity of HIV-infected individuals. Nevertheless long-term toxicity and appearance of viral resistance hampers the prolonged effectiveness of combination therapy, requiring a continuous input of drugs to replace those utilized in combination regimens. We here investigated the anti-HIV activity of novel derivatives of the suradista chemical class., Methods: Compounds were tested on acute HIV-1 infection of activated peripheral blood mononuclear cells. HIV production was monitored by enzyme-linked immunosorbent assay measuring the protein p24 released in culture supernatants. Fusion assays were carried out to study the mechanism of action of these compounds. A modified version of a previously established recombinant vaccinia virus-based assay was used measuring activation of a reporter gene upon fusion of two distinct cell populations. Flow cytometry was performed in competition assays for the binding of several antibodies targeting different sites of the viral envelope glycoprotein gp120, or the receptor CD4, or the coreceptors CXCR4 and CCR5., Results: Four compounds inhibited replication of a prototypic R5 (BaL) and X4 (IIIB) laboratory-adapted HIV-1 strain at low micromolar concentrations, in the absence of cytotoxicity. Approximately a ten fold greater activity was achieved against the X4 as compared to the R5 strain. The compounds blocked X4 and R5 HIV-1 fusion, a step of viral entry. This activity appeared specific for HIV-1, as entry of human herpesvirus 6 (HHV-6) and influenza virus was not substantially affected. Further investigation of the inhibitory mechanism revealed that these new molecules target the viral envelope, rather than the coreceptors, as previously shown for a congener of the same class characterized by a long plasmatic half-life. Indeed ND-4043, the most active compound, specifically competed with binding of monoclonal antibodies against the CD4-binding site (CD4-BS) and coreceptor-binding site (CoR-BS) of gp120. These compounds displayed broad anti-HIV activity, as they inhibited various primary R5, X4 and, importantly, dualtropic R5X4 HIV-1 isolates. Of the four derivatives tested, the dimeric compounds were consistently more potent than the monomeric ones., Conclusions: Given their unique features, these molecules represent promising candidates for further development and exploitation as anti-HIV therapeutics.
- Published
- 2015
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42. Tyrosine sulfation in the second variable loop (V2) of HIV-1 gp120 stabilizes V2-V3 interaction and modulates neutralization sensitivity.
- Author
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Cimbro R, Gallant TR, Dolan MA, Guzzo C, Zhang P, Lin Y, Miao H, Van Ryk D, Arthos J, Gorshkova I, Brown PH, Hurt DE, and Lusso P
- Subjects
- Blotting, Western, Flow Cytometry, HEK293 Cells, HIV Envelope Protein gp120 genetics, Humans, Neutralization Tests, Surface Plasmon Resonance, Tyrosine metabolism, HIV Envelope Protein gp120 chemistry, HIV Envelope Protein gp120 metabolism, HIV-1 genetics, HIV-1 immunology, Protein Conformation, Tyrosine analogs & derivatives
- Abstract
Elicitation of broadly neutralizing antibodies is essential for the development of a protective vaccine against HIV-1. However, the native HIV-1 envelope adopts a protected conformation that conceals highly conserved sites of vulnerability from antibody recognition. Although high-definition structures of the monomeric core of the envelope glycoprotein subunit gp120 and, more recently, of a stabilized soluble gp140 trimer have been solved, fundamental aspects related to the conformation and function of the native envelope remain unresolved. Here, we show that the conserved central region of the second variable loop (V2) of gp120 contains sulfated tyrosines (Tys173 and Tys177) that in the CD4-unbound prefusion state mediate intramolecular interaction between V2 and the conserved base of the third variable loop (V3), functionally mimicking sulfated tyrosines in CCR5 and anti-coreceptor-binding-site antibodies such as 412d. Recombinant gp120 expressed in continuous cell lines displays low constitutive levels of V2 tyrosine sulfation, which can be enhanced markedly by overexpression of the tyrosyl sulfotransferase TPST2. In contrast, virion-associated gp120 produced by primary CD4(+) T cells is inherently highly sulfated. Consistent with a functional role of the V2 sulfotyrosines, enhancement of tyrosine sulfation decreased binding and neutralization of HIV-1 BaL by monomeric soluble CD4, 412d, and anti-V3 antibodies and increased recognition by the trimer-preferring antibodies PG9, PG16, CH01, and PGT145. Conversely, inhibition of tyrosine sulfation increased sensitivity to soluble CD4, 412d, and anti-V3 antibodies and diminished recognition by trimer-preferring antibodies. These results identify the sulfotyrosine-mediated V2-V3 interaction as a critical constraint that stabilizes the native HIV-1 envelope trimer and modulates its sensitivity to neutralization.
- Published
- 2014
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43. Subcutaneous golimumab maintains clinical response in patients with moderate-to-severe ulcerative colitis.
- Author
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Sandborn WJ, Feagan BG, Marano C, Zhang H, Strauss R, Johanns J, Adedokun OJ, Guzzo C, Colombel JF, Reinisch W, Gibson PR, Collins J, Järnerot G, and Rutgeerts P
- Subjects
- Adrenal Cortex Hormones therapeutic use, Adult, Aminosalicylic Acids therapeutic use, Double-Blind Method, Drug Therapy, Combination, Female, Humans, Injections, Subcutaneous, Maintenance Chemotherapy methods, Male, Mercaptopurine therapeutic use, Methotrexate therapeutic use, Middle Aged, Severity of Illness Index, Treatment Outcome, Anti-Inflammatory Agents therapeutic use, Antibodies, Monoclonal therapeutic use, Colitis, Ulcerative drug therapy, Immunosuppressive Agents therapeutic use, Tumor Necrosis Factor-alpha antagonists & inhibitors
- Abstract
Background & Aims: Subcutaneous golimumab, a fully human monoclonal antibody to tumor necrosis factor-α (TNFα), was evaluated as maintenance therapy in TNFα antagonist-naive adults with moderate-to-severe active ulcerative colitis, despite conventional therapy, who responded to golimumab induction therapy., Methods: We performed a phase 3, double-blind trial of patients who completed golimumab induction trials (Program of Ulcerative Colitis Research Studies Utilizing an Investigational Treatment, eg, PURSUIT). Patients who responded to induction therapy with golimumab (n = 464) were assigned randomly to groups given placebo or injections of 50 or 100 mg golimumab every 4 weeks through week 52. Patients who responded to placebo in the induction study continued to receive placebo. Nonresponders in the induction study received 100 mg golimumab. The primary end point was clinical response maintained through week 54; secondary end points included clinical remission and mucosal healing at both weeks 30 and 54., Results: Clinical response was maintained through week 54 in 47.0% of patients receiving 50 mg golimumab, 49.7% of patients receiving 100 mg golimumab, and 31.2% of patients receiving placebo (P = .010 and P < .001, respectively). At weeks 30 and 54, a higher percentage of patients who received 100 mg golimumab were in clinical remission and had mucosal healing (27.8% and 42.4%) than patients given placebo (15.6% and 26.6%; P = .004 and P = .002, respectively) or 50 mg golimumab (23.2% and 41.7%, respectively). Percentages of serious adverse events were 7.7%, 8.4%, and 14.3% among patients given placebo, 50 mg, or 100 mg golimumab, respectively; percentages of serious infections were 1.9%, 3.2%, and 3.2%, respectively. Among all patients given golimumab in the study, 3 died (from sepsis, tuberculosis, and cardiac failure, all in patients who received 100 mg golimumab) and 4 developed active tuberculosis., Conclusions: Golimumab (50 mg or 100 mg) maintained clinical response through week 54 in patients who responded to induction therapy with golimumab and had moderate-to-severe active ulcerative colitis; patients who received 100 mg golimumab had clinical remission and mucosal healing at weeks 30 and 54. Safety was consistent with that reported for other TNFα antagonists and golimumab in other approved indications. ClinicalTrials.gov number: NCT00488631., (Copyright © 2014 AGA Institute. Published by Elsevier Inc. All rights reserved.)
- Published
- 2014
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44. Subcutaneous golimumab induces clinical response and remission in patients with moderate-to-severe ulcerative colitis.
- Author
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Sandborn WJ, Feagan BG, Marano C, Zhang H, Strauss R, Johanns J, Adedokun OJ, Guzzo C, Colombel JF, Reinisch W, Gibson PR, Collins J, Järnerot G, Hibi T, and Rutgeerts P
- Subjects
- Adrenal Cortex Hormones therapeutic use, Adult, Aminosalicylic Acids therapeutic use, Azathioprine therapeutic use, Dose-Response Relationship, Drug, Double-Blind Method, Drug Therapy, Combination, Female, Humans, Injections, Subcutaneous, Male, Mercaptopurine therapeutic use, Methotrexate therapeutic use, Middle Aged, Quality of Life, Remission Induction methods, Treatment Outcome, Anti-Inflammatory Agents therapeutic use, Antibodies, Monoclonal therapeutic use, Colitis, Ulcerative drug therapy, Immunosuppressive Agents therapeutic use, Tumor Necrosis Factor-alpha antagonists & inhibitors
- Abstract
Background & Aims: Little is known about the efficacy of golimumab, a fully human monoclonal antibody to tumor necrosis factor (TNF) -α, for treatment of ulcerative colitis (UC). We evaluated subcutaneous golimumab induction therapy in TNF-α antagonist-naïve patients with moderate-to-severe UC despite conventional treatment., Methods: We integrated double-blind phase 2 dose-finding and phase 3 dose-confirmation trials in a study of 1064 adults with UC (Mayo score: 6-12; endoscopic subscore ≥ 2; 774 patients in phase 3). Patients were randomly assigned to groups given golimumab doses of 100 mg and then 50 mg (phase 2 only), 200 mg and then 100 mg, or 400 mg and then 200 mg, 2 weeks apart. The phase 3 primary end point was week-6 clinical response. Secondary end points included week-6 clinical remission, mucosal healing, and Inflammatory Bowel Disease Questionnaire (IBDQ) score change., Results: In phase 2, median changes from baseline in the Mayo score were -1.0, -3.0, -2.0, and -3.0, in the groups given placebo, 100 mg/50 mg, 200/100 mg, and 400/200 mg golimumab, respectively. In phase 3, rates of clinical response at week 6 were 51.0% and 54.9% among patients given 200 mg/100 mg and 400 mg/200 mg golimumab, respectively, vs 30.3% among those given placebo (both, P ≤ .0001). Rates of clinical remission and mucosal healing and mean changes in IBDQ scores were significantly greater in both golimumab groups vs the placebo group (P ≤ .0014, all comparisons). Rates of serious adverse events were 6.1% and 3.0%, and rates of serious infection were 1.8% and 0.5%, in the placebo and golimumab groups, respectively. One patient in the 400 mg/200 mg group died as a result of surgical complications of an ischiorectal abscess., Conclusions: Treatment with subcutaneous golimumab induces clinical response, remission, and mucosal healing, and increases quality of life in larger percentages of patients with active UC than placebo. ClinicalTrials.gov Number: NCT00487539., (Copyright © 2014 AGA Institute. Published by Elsevier Inc. All rights reserved.)
- Published
- 2014
- Full Text
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45. Phenformin enhances the therapeutic benefit of BRAF(V600E) inhibition in melanoma.
- Author
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Yuan P, Ito K, Perez-Lorenzo R, Del Guzzo C, Lee JH, Shen CH, Bosenberg MW, McMahon M, Cantley LC, and Zheng B
- Subjects
- Analysis of Variance, Animals, Antineoplastic Combined Chemotherapy Protocols pharmacology, Apoptosis drug effects, Cell Proliferation drug effects, DNA-Binding Proteins metabolism, Immunohistochemistry, Indoles administration & dosage, Jumonji Domain-Containing Histone Demethylases metabolism, Melanoma genetics, Mice, Mutation, Missense genetics, Phenformin administration & dosage, Proto-Oncogene Proteins B-raf genetics, Sulfonamides administration & dosage, Indoles pharmacology, Melanoma drug therapy, Phenformin pharmacology, Proto-Oncogene Proteins B-raf antagonists & inhibitors, Sulfonamides pharmacology
- Abstract
Biguanides, such as the diabetes therapeutics metformin and phenformin, have demonstrated antitumor activity both in vitro and in vivo. The energy-sensing AMP-activated protein kinase (AMPK) is known to be a major cellular target of biguanides. Based on our discovery of cross-talk between the AMPK and v-Raf murine sarcoma viral oncogene homolog B1 (BRAF) signaling pathways, we investigated the antitumor effects of combining phenformin with a BRAF inhibitor PLX4720 on the proliferation of BRAF-mutated melanoma cells in vitro and on BRAF-driven tumor growth in vivo. Cotreatment of BRAF-mutated melanoma cell lines with phenformin and PLX4720 resulted in synergistic inhibition of cell viability, compared with the effects of the single agent alone. Moreover, treatment with phenformin significantly delayed the development of resistance to PLX4720 in cultured melanoma cells. Biochemical analyses showed that phenformin and PLX4720 exerted cooperative effects on inhibiting mTOR signaling and inducing apoptosis. Noticeably, phenformin selectively targeted subpopulations of cells expressing JARID1B, a marker for slow cycling melanoma cells, whereas PLX4720 selectively targeted JARID1B-negative cells. Finally, in contrast to their use as single agents, the combination of phenformin and PLX4720 induced tumor regression in both nude mice bearing melanoma xenografts and in a genetically engineered BRAF(V600E)/PTEN(null)-driven mouse model of melanoma. These results strongly suggest that significant therapeutic advantage may be achieved by combining AMPK activators such as phenformin with BRAF inhbitors for the treatment of melanoma.
- Published
- 2013
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46. Two RND proteins involved in heavy metal efflux in Caulobacter crescentus belong to separate clusters within proteobacteria.
- Author
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Valencia EY, Braz VS, Guzzo C, and Marques MV
- Subjects
- Biological Transport, Active, Cluster Analysis, Evolution, Molecular, Gene Deletion, Gene Expression Profiling, Phylogeny, Sequence Homology, Amino Acid, Caulobacter crescentus genetics, Caulobacter crescentus metabolism, Membrane Transport Proteins genetics, Membrane Transport Proteins metabolism, Metals, Heavy metabolism, Multigene Family
- Abstract
Background: Heavy metal Resistance-Nodulation-Division (HME-RND) efflux systems help Gram-negative bacteria to keep the intracellular homeostasis under high metal concentrations. These proteins constitute the cytoplasmic membrane channel of the tripartite RND transport systems. Caulobacter crescentus NA1000 possess two HME-RND proteins, and the aim of this work was to determine their involvement in the response to cadmium, zinc, cobalt and nickel, and to analyze the phylogenetic distribution and characteristic signatures of orthologs of these two proteins., Results: Expression assays of the czrCBA operon showed significant induction in the presence of cadmium and zinc, and moderate induction by cobalt and nickel. The nczCBA operon is highly induced in the presence of nickel and cobalt, moderately induced by zinc and not induced by cadmium. Analysis of the resistance phenotype of mutant strains showed that the ΔczrA strain is highly sensitive to cadmium, zinc and cobalt, but resistant to nickel. The ΔnczA strain and the double mutant strain showed reduced growth in the presence of all metals tested. Phylogenetic analysis of the C. crescentus HME-RND proteins showed that CzrA-like proteins, in contrast to those similar to NczA, are almost exclusively found in the Alphaproteobacteria group, and the characteristic protein signatures of each group were highlighted., Conclusions: The czrCBA efflux system is involved mainly in response to cadmium and zinc with a secondary role in response to cobalt. The nczCBA efflux system is involved mainly in response to nickel and cobalt, with a secondary role in response to cadmium and zinc. CzrA belongs to the HME2 subfamily, which is almost exclusively found in the Alphaproteobacteria group, as shown by phylogenetic analysis. NczA belongs to the HME1 subfamily which is more widespread among diverse Proteobacteria groups. Each of these subfamilies present distinctive amino acid signatures.
- Published
- 2013
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47. The CD8-derived chemokine XCL1/lymphotactin is a conformation-dependent, broad-spectrum inhibitor of HIV-1.
- Author
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Guzzo C, Fox J, Lin Y, Miao H, Cimbro R, Volkman BF, Fauci AS, and Lusso P
- Subjects
- CD4 Antigens metabolism, Cells, Cultured, Chemokines, C chemistry, Chemokines, C pharmacology, HIV Envelope Protein gp120 metabolism, Humans, Protein Binding, Protein Conformation, Protein Folding, Receptors, CCR5 metabolism, Receptors, CXCR4 metabolism, Structure-Activity Relationship, Virus Attachment drug effects, Virus Internalization drug effects, CD8-Positive T-Lymphocytes immunology, Chemokines, C immunology, HIV-1 physiology
- Abstract
CD8+ T cells play a key role in the in vivo control of HIV-1 replication via their cytolytic activity as well as their ability to secrete non-lytic soluble suppressive factors. Although the chemokines that naturally bind CCR5 (CCL3/MIP-1α, CCL4/MIP- 1β, CCL5/RANTES) are major components of the CD8-derived anti-HIV activity, evidence indicates the existence of additional, still undefined, CD8-derived HIV-suppressive factors. Here, we report the characterization of a novel anti-HIV chemokine, XCL1/lymphotactin, a member of the C-chemokine family that is produced primarily by activated CD8+ T cells and behaves as a metamorphic protein, interconverting between two structurally distinct conformations (classic and alternative). We found that XCL1 inhibits a broad spectrum of HIV-1 isolates, irrespective of their coreceptor-usage phenotype. Experiments with stabilized variants of XCL1 demonstrated that HIV-1 inhibition requires access to the alternative, all-β conformation, which interacts with proteoglycans but does not bind/activate the specific XCR1 receptor, while the classic XCL1 conformation is inactive. HIV-1 inhibition by XCL1 was shown to occur at an early stage of infection, via blockade of viral attachment and entry into host cells. Analogous to the recently described anti-HIV effect of the CXC chemokine CXCL4/PF4, XCL1-mediated inhibition is associated with direct interaction of the chemokine with the HIV-1 envelope. These results may open new perspectives for understanding the mechanisms of HIV-1 control and reveal new molecular targets for the design of effective therapeutic and preventive strategies against HIV-1.
- Published
- 2013
- Full Text
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48. Induction and maintenance therapy with infliximab for children with moderate to severe ulcerative colitis.
- Author
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Hyams J, Damaraju L, Blank M, Johanns J, Guzzo C, Winter HS, Kugathasan S, Cohen S, Markowitz J, Escher JC, Veereman-Wauters G, Crandall W, Baldassano R, and Griffiths A
- Subjects
- Adolescent, Antibodies, Monoclonal adverse effects, Child, Colitis, Ulcerative pathology, Female, Humans, Induction Chemotherapy methods, Infliximab, Maintenance Chemotherapy methods, Male, Severity of Illness Index, Treatment Outcome, Anti-Inflammatory Agents administration & dosage, Antibodies, Monoclonal administration & dosage, Colitis, Ulcerative drug therapy, Gastrointestinal Agents administration & dosage
- Abstract
Background & Aims: We evaluated the efficacy and safety of infliximab for inducing and maintaining benefit in children with moderately to severely active ulcerative colitis (UC)., Methods: Patients (6-17 years old) who had active UC (Mayo scores of 6-12; endoscopic subscores ≥ 2) and had not responded to or tolerated conventional treatment were given 5 mg/kg infliximab at weeks 0, 2, and 6. The primary end point was response at week 8 (decreases in Mayo scores ≥ 30% and ≥ 3 points and decreases in rectal bleeding subscores of ≥ 1 or an absolute subscore of ≤ 1). At week 8, only responders were randomly assigned to groups given infliximab every 8 or 12 weeks (q8w or q12w) and followed through week 54. Maintenance end points included pediatric UC activity index scores <10 points, defined as remission., Results: At week 8, infliximab induced a response in 73.3% of patients (44 of 60) (95% confidence interval, 62.1%-84.5%; a positive result was defined by 95% confidence interval lower limit >40%). Among responders, twice as many were in remission at week 54 after q8w (8 of 21, 38.1%) than q12w (4 of 22, 18.2%; P = .146) therapy. Assuming the q8w remission rate for responders, the overall remission rate at week 54 would be 28.6%. Serious adverse events and infusion reactions occurred in similar proportions in the q8w and q12w groups. No deaths, malignancies, opportunistic infections, tuberculosis, or delayed hypersensitivity reactions were reported., Conclusions: Infliximab was safe and effective, inducing a response at week 8 in 73.3% of pediatric patients with moderate to severely active UC who did not respond to conventional therapy. The overall remission rate at week 54 for all enrolled patients was 28.6%, assuming the more effective q8w remission rate., (Copyright © 2012 AGA Institute. Published by Elsevier Inc. All rights reserved.)
- Published
- 2012
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49. IL-27-induced gene expression is downregulated in HIV-infected subjects.
- Author
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Guzzo C, Hopman WM, Che Mat NF, Wobeser W, and Gee K
- Subjects
- Down-Regulation, Enzyme-Linked Immunosorbent Assay methods, Flow Cytometry methods, Humans, Inflammation, Leukocytes, Mononuclear cytology, Monocytes cytology, Monocytes virology, Real-Time Polymerase Chain Reaction methods, env Gene Products, Human Immunodeficiency Virus metabolism, Gene Expression Regulation, Viral, HIV Infections metabolism, HIV Infections virology, Interleukins biosynthesis
- Abstract
Objective: To characterize the effect of HIV infection on IL-27-induced gene expression., Design: During HIV infection, cytokine expression and function become deregulated. IL-27 is an important modulator of inflammatory responses. Interestingly, IL-27 can inhibit HIV replication in T cells and monocytes, implicating IL-27 as a potential adjunct to anti-viral treatment. Our previous work demonstrated that circulating HIV may suppress IL-27 expression, therefore, this study, in continuation of our previous work, aimed to understand how HIV affects expression levels of the IL-27 receptor and downstream functions of IL-27., Methods: Peripheral blood mononuclear cells (PBMC) were isolated from whole blood of HIV negative and HIV positive (viremic) individuals to assess IL-27-induced gene expression by flow cytometry and ELISA. PBMC were also processed for monocyte enrichment to assess IL-27 receptor expression by flow cytometry and real-time PCR., Results: Expression of the IL-27 receptor subunit, gp130, was upregulated in response to IL-27 in HIV negative individuals, however, in HIV positive individuals, this IL-27 response was diminished. Furthermore, we observed downregulation of IL-27-induced IL-6, TNF-α, and IL-10 expression in HIV positive subjects., Conclusion: In HIV infection, IL-27-induced gene expression was impaired, indicating HIV-mediated dysregulation of IL-27 functions occurs during HIV infection. This study provides evidence for new viral pathogenic mechanisms contributing to the widespread impairment of immune responses observed in HIV pathogenesis.
- Published
- 2012
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50. IL-27 increases BST-2 expression in human monocytes and T cells independently of type I IFN.
- Author
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Guzzo C, Jung M, Graveline A, Banfield BW, and Gee K
- Subjects
- APOBEC Deaminases, Cell Line, Cytidine Deaminase, Flow Cytometry, GPI-Linked Proteins genetics, Humans, RNA, Messenger genetics, Signal Transduction, Antigens, CD genetics, Cytosine Deaminase metabolism, Interferon Type I metabolism, Interleukin-17 physiology, Monocytes metabolism, T-Lymphocytes metabolism
- Abstract
IL-27 modulates inflammatory responses by influencing cytokine secretion and CD4 T cell differentiation. Recently, IL-27 was demonstrated to inhibit HIV replication by inducing type I interferon (IFN) expression and subsequent IFN-dependent expression of apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like (APOBEC)-3 family members, a group of antiviral cytidine deaminases. To characterize other anti-viral genes modulated by IL-27, we examined another IFN-responsive gene: tetherin/bone marrow stromal cell antigen 2 (BST-2). Our study shows that IL-27 can directly induce BST-2 expression, independently of an intermediary type I IFN response. Quantitative RT-PCR analysis demonstrated IL-27-induced BST-2 mRNA expression as early as 2h after exposure of cells to IL-27. In the presence of the type I IFN-neutralizing protein, B18R, IL-27-induced BST-2 expression was maintained, demonstrating that IFN is not an intermediary in IL-27-induced BST-2. Taken together, our findings identify a novel function of IL-27 as a direct stimulator of BST-2 expression.
- Published
- 2012
- Full Text
- View/download PDF
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