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Rachelle Mendoza,1 Michael Silver,2,3 Alejandro R Zuretti,1,4 Manan Christian,4 Ballabh Das,5 Allen J Norin,5 Patrick Borgen,6 Jenny Libien,1,4 Martin H Bluth4 1Department of Pathology, SUNY Downstate Health Sciences University, Brooklyn, NY, USA; 2School of Public Health, SUNY Downstate Health Sciences University, Brooklyn, NY, USA; 3Department of Medicine, Maimonides Medical Center, Brooklyn, NY, USA; 4Department of Pathology, Maimonides Medical Center, Brooklyn, NY, USA; 5Department of Medicine, SUNY Downstate Health Sciences University, Brooklyn, NY, USA; 6Department of Surgery, Maimonides Medical Center, Brooklyn, NY, USACorrespondence: Martin H BluthBlood Transfusion and Donor Services, Clinical Laboratories, Translational Research (Pathology), Maimonides Medical Center, 4802 10 th Avenue, Brooklyn, NY, 11219, USAEmail MBluth@maimonidesmed.orgBackground: We investigated the utility of an automated chemiluminescent SARS-CoV-2 IgG antibody assay platform in quantifying the amount of binding antibodies present in donated convalescent plasma.Methods: A total of 179 convalescent plasma units were analyzed for the presence of SARS-CoV-2 IgG antibodies using the Beckman-Coulter chemiluminescent immunoassay (CLIA) platform. The equipment-derived numerical values (S/Co ratio) were recorded. Aliquots from the same units were subjected to enzyme-linked immunosorbent assay (ELISA) that detects IgG antibodies against the receptor-binding domain (RBD) of the SARS-CoV-2 S1 protein. The relationship between ELISA titers and CLIA S/Co values was analyzed using linear regression and receiver operating characteristics (ROC) curve.Results: Twenty-one samples (11.7%) had S/Co values of less than 1.0 and were deemed negative for antibodies and convalescent plasma had S/Co values between > 1.0 and 5.0 (70/179, 39.1%). Fifteen units (8.4%) had negative ELISA titer. The majority of the units (95/179. 53.1%) had titers ≥ 1:1024. The sensitivities of ELISA to CLIA were comparable (90.5% vs 88.3%, respectively; p=0.18). There was positive linear correlation between CLIA S/Co values and ELISA IgG titer (Rho = 0.75; Spearman’s rank = 0.82, p-value = < 0.0001). The agreement between the two methods was fair, with a κ index of 0.2741. Using the ROC analysis, we identified a CLIA S/Co cutoff value of 8.2, which gives a sensitivity of 90% and a specificity of 82% in predicting a titer dilution of ≥ 1:1024.Conclusion: The utility of automated antibody detection systems can be extended from simply a screening method to a semi-quantitative and quantitative functional antibody analysis. CLIA S/Co values can be used to reliably estimate the ELISA antibody titer. Incorporation of chemiluminescent-based methods can provide rapid, cost-effective means of identifying anti-SARS-CoV-2 antibody titers in donated plasma for use in the treatment of COVID-19 infection.Keywords: COVID-19, ELISA, chemiluminescence, antibody, titers

Virtually all the red blood cell and platelet antigen systems have been characterized at the molecular level. Highly reliable methods for red blood cell and platelet antigen genotyping are now available. Genotyping is a useful adjunct to traditional serology and can help resolve complex serologic problems. Although red blood cell and platelet phenotypes can be inferred from genotype, knowledge of the molecular basis is essential for accurate assignment. Genotyping of blood donors is an effective method of identifying antigen-negative and/or particularly rare donors. Cell-free DNA analysis provides a promising noninvasive method of assessing fetal genotypes of blood group alloantigens.

Pancreatic neoplasms, including ductal adenocarcinoma, solid pseudopapillary neoplasm, pancreatic endocrine neoplasms, acinar cell carcinoma, and pancreatoblastoma, are associated with different genetic abnormalities. Hepatic adenomas with beta-catenin exon 3 mutation are associated with a high risk of malignancy. Hepatic adenoma with arginosuccinate synthetase 1 expression or sonic hedgehog mutations are associated with a risk of bleeding. Hepatocellular carcinoma and choangiocarcinoma display heterogeneity at both morphologic and molecular levels Cholangiocellular carcinoma is most commonly associated with IDH 1/2 mutations.

Precision medicine applies primarily to pharmacokinetics in toxicology. Mastering hepatic metabolism through an understanding of the genetics behind Phase I, or oxidation/reduction and some Phase II, or conjugation, enhances the scientific and clinical application of common drug toxicology. This review includes basic hepatic metabolism, the common substrates, inducers and inhibitors of cytochrome P450 along with genetic variants behind the enzymes. Detailed metabolism of commonly measured opioids in clinical practice is provided. Finally, evidence based research and clinical correlations conclude that knowledge of inducers and inhibitors, in conjunction with genetic variations, are integral components for applied precision medicine in toxicology.

Respiratory syncytial virus (RSV) causes lower respiratory tract disease in infants and young children, and is a public health concern, as is the increase in pediatric asthma. Respiratory viral infections may trigger asthma exacerbations. However, it remains unknown whether RSV infection may have a specific association with asthma. Total serum IgE, and IgE- and IgG-anti-RSV Ab responses were studied in older asthmatic compared with non-asthmatic children (M/F, mean age: 14) (N=30, N=43, respectively). We found: (1) total serum IgE was higher in asthmatic compared with non-asthmatics (P

The use of probiotics has gained considerable popularity in the marketplace in the US and has been steadily increasing due to consumer interest in potential treatment of various diseases, which may be due to modulation of immune responses. The aim of this review is to present information from representative studies regarding some of the possible applications and clinical effects of probiotic use in diseases of altered immunoglobulin (IgE) regulation (allergic rhinitis (AR), asthma, atopic dermatitis (AD) and food sensitization). Reports in humans are sparse or controversial; there is currently little reliable scientific data that supports the theory that there exists a cause–effect relationship between taking probiotics and alleviation of allergic disease. Unfortunately, these findings are too variable to allow substantial conclusions as to the efficacy and effectiveness of probiotic use in these disease states.

Rationale The role of peripheral blood progenitor cell mobilization on Immunoglobulin E (IgE) responses has not been studied. Methods Distributions of blood lymphocytes (CD4+, CD8+, CD8+CD60+, CD19+, CD23+, CD16/56+, CD25, CD45RA+, CD45RO+, CD34+), and levels of serum immunoglobulins (IgM, IgG, IgA, IgE) were studied in an allergic asthmatic serum IgE+ (181 IU/mL) adult (m/45 y/o) donor undergoing routine stem cell mobilization protocol (American Society of Hematology) before (day-30), during (day 4), and after (1 wk post last dose) filgrastim (subcutaneous, 480 mcg, 2qd) treatment (flow cytometry, nephelometry, UniCAP Total IgE Fluoro enzyme immunoassay). Results On day 4 of filgrastim treatment, numbers of CD8+CD60+T cells and CD23+ blood cells dramatically increased (98% and 240% respectively) compared with pre treatment. In contrast on day 4 of treatment, serum IgE levels decreased (>50%) compared with pre treatment. CD8+CD60+T cells and CD23+ blood cells and serum IgE levels approached pre-treatment levels at 1 week post treatment. Conclusions Filgrastim treatment transiently increases numbers of CD8+CD60+T and CD23+ expressing cells, which are known to regulate human IgE responses, while also transiently suppressing ongoing IgE responses. These results suggest that filgrastim affects IgE related responses, and may be useful in modulating allergic responses.

Bronchial asthma is exacerbated by Mycoplasma pneumoniae-induced upper respiratory tract infections (URTIs) in children. Specific IgM and IgG isotypes are involved in the immune response to M. pneumoniae, but little is known about the role of specific IgE antibodies against M. pneumoniae in asthma.To investigate the role of IgM-, IgG- and IgE-specific antibody responses to M. pneumoniae in children with persistent asthma in relationship to history of URTI within the past 6 months.Total or specific anti-M. pneumoniae IgM, IgG and IgE antibody responses were studied in stable asthmatic pediatric patients (M. pneumoniae positive and negative) without current exacerbation and nonasthmatic controls (N = 23 and 13, respectively) (UniCAP total IgE Fluoroenzymeimmunoassay, enzyme-linked immunosorbent assay).Values of specific IgM correlated with specific IgG (Spearman correlation, rho = 0.61, P0.0001) but not with specific IgE anti-M. pneumoniae antibodies (AMA) in asthmatic subjects compared with nonasthmatic controls. However, concentrations of specific IgG correlated with specific IgE AMA (rho = 0.49, P = 0.0017). Asthmatic subjects had higher levels of specific IgM AMA levels compared with nonasthmatics (median [interquartile range]: 0.57 [1.00] versus 0.21 [0.19]; Kruskal-Wallis test, P = 0.0008). In addition, IgM positivity was significantly higher in asthmatic compared with nonasthmatic subjects (39.1% versus 0.0%; Fisher's exact test, P = 0.01). These results were independent of URTI history in the past 6 months, which was not associated with higher IgM, IgG or IgE AMA levels compared with no URTI history (P = 0.25-0.64).Increased specific IgM anti-M. pneumoniae responses may indicate an important role for M. pneumoniae infection in asthma.

Tamar A Smith-Norowitz,1 Melanie Kusonruksa,2 Darrin Wong,2 Moshe M Norowitz,2 Rauno Joks,3 Helen G Durkin,2 Martin H Bluth41Department of Pediatrics, 2Department of Pathology, 3Department of Medicine, Center for Allergy and Asthma Research, SUNY Downstate Medical Center, Brooklyn, New York, NY; 4Department of Pathology, Wayne State University School of Medicine, Detroit, MI, USABackground and methods: The role of immunoglobulin (Ig) E in immunity against influenza A H1N1 has not been studied. Total serum IgE and specific IgE and IgG anti-H1N1 virus responses were studied in children and adults (n = 2) who received influenza virus vaccination (Flumist® or Fluzone® ) in autumn 2008 and 2009, and then subsequently became infected with the H1N1 virus in spring 2009. Twelve months after infection, antibodies in their serum were compared with those in the serum of subjects who were either vaccinated but not infected (n = 4) or nonvaccinated and noninfected subjects (n = 2), using UniCAP total IgE fluoroenzyme immunoassay, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and Western blotting. Band sizes for the influenza virus (58, 56, 40, 30, 25, and 17 kDa) and H1N1 viral proteins (58, 56, 25, and 17 kDa) were determined, using sodium dodecyl sulfate polyacrylamide gel electrophoresis and Coomassie brilliant blue.Results: We found that the serum of vaccinated and subsequently infected children and adults contained IgE and IgG antibodies to both H1N1 and influenza virus, with a strong IgE and IgG band intensity at 56 kDa. Interestingly, in subjects who were vaccinated but not infected, band intensity at 56 kDa was lowered by approximately two-fold. Serum of nonvaccinated and noninfected subjects had no detectable IgE or IgG antibodies to influenza virus or H1N1.Conclusion: This is the first description of IgE anti-influenza A H1N1 antibodies in human serum and the first demonstration of their long-term persistence. The decreased intensity of the 56 kDa band in vaccinated noninfected subjects compared with vaccinated infected subjects suggests augmented IgE and IgG antibody responses to influenza A H1N1.Keywords: influenza A H1N1, immunoglobulin E, vaccination

The nexus of chemotherapeutic intervention and the immunomodulation of IgE-related phenomena are not well understood. The relationship bears importance in better understanding the causal and/or resultant effects of one on the other and their collective role in the management and sequelae of the cancer patient. This review discusses the relationship of chemotherapy on immunoglobulins with a focus on IgE and other related biological processes including hypersensitivity reactions and proposes models toward effective management of the cancer patient in this regard.

The production of IgE specific to different viruses (HIV-1, Parvovirus B19, Parainfluenza virus, Varicella Zoster Virus), and the ability of IgE anti-HIV-1 to suppress HIV-1 production in vitro, strongly suggest an important role for IgE and/or anti viral specific IgE in viral pathogenesis. Nevertheless, the presence and persistence of IgE anti-Influenza virus antibodies has not been studied. Total serum IgE and specific IgE and IgG anti-Influenza virus antibodies were studied in children (N = 3) (m/f 14-16 y/o) and adults (N = 3) (m/f, 41-49 y/o) 2-20 months after vaccination with Influenza virus (Flumist(®) or Fluzone(®)), as well as in non-vaccinated children (N = 2). (UniCAP total IgE Fluoroenzymeimmunoassay, ELISA, Immunoblot). We found that serum of vaccinated children and adults contained IgE and IgG anti-Influenza virus antibodies approaching two years post vaccination. Non-vaccinated children did not make either IgE or IgG anti-Influenza antibodies. Similar levels of IL-2, IFN-γ, IL-4, and IL-10 cytokines were detected in serum of vaccinated compared with non vaccinated subjects (p0.05), as well as between vaccinated adults compared with vaccinated children and non vaccinated subjects (p0.05). Vaccinated children and adults continue to produce IgE anti-Influenza virus antibodies long term post vaccination. The long term production of IgE anti-Influenza virus antibodies induced by vaccination may contribute to protective immunity against Influenza.

Rationale The role of the immune response to caterpillar exposure is not well described. This case study is the first to report a patient who presented with an allergic reaction after exposure to the larvae of the sycamore tussock moth, Halysidota harrisii Walsh, 1864. Methods Blood was collected from an allergic asthmatic adult (m/42 y/o) at 2 hrs – 2 wks after contact urticaria with associated dyspnea after exposure to the larvae of the sycamore tussock moth, Halysidota harrisii Walsh, 1864. Distributions of blood lymphocytes (CD4+, CD8+, CD8+CD60+, CD19+, CD23+, CD16/56+, CD25, CD45RA+, CD45RO+), monocytes (CD1d+), levels of serum immunoglobulins (IgM, IgG, IgA, IgE), and cytokines (IFN-γ, IL-4, TNF-α) were studied (flow cytometry, nephelometry, UniCAP Total IgE Fluoroenzymeimmunoassay, cytokine ELISA, clinical toxicology). Results Numbers of CD4+ T cells, CD25+ cells, CD19+ B cells, and CD1d+ monocytes decreased (22, 27, 33, 20%, respectively) one week post reaction, CD45RA+ naïve T cells decreased at 36 hours (21%),while CD8+CD60+ T cells and CD23+ cells decreased 48 hrs (33, 74%, respectively) post reaction. In contrast, numbers of CD16/56+ NK precursor cells increased (60%) 12 hrs, then decreased (65%) 48 hrs post reaction; other lymphocyte subsets were unaffected. Serum IgM, IgG and IgA were within normal range; however, serum IgE demonstrated a bimodal elevation at 2 hrs (15%) and one week post reaction. Levels of IFN-γ, IL-4, and TNF-α were not detected in serum pre-exposure (

The role of the immune response in autoimmune hepatitis has not been studied before and after prednisone and azathioprine treatment. Distributions of blood lymphocytes (CD4+, CD8+, CD19+, CD23+, CD16/56+), levels of serum immunoglobulins (IgM, IgG, IgE, IgA) and cytokines (IFN-gamma, IL-4, IL-12, TNFalpha ) were studied in a child (f/14 y/o) with autoimmune hepatitis before and after prednisone (20 mg/d) and azathioprine (50 mg/d) treatment (nephelometry, UniCAP Total IgE Fluoroenzymeimmunoassay, flow cytometry, cytokine ELISA). Patient was studied for 0-2.5 yrs; treatment was initiated 12 weeks post diagnosis. Numbers of CD4+ T cells increased (50%), while CD19+ and CD23+ cells decreased (50%) post treatment; other lymphocyte subsets were unaffected by treatment. Serum IgG and IgE levels decreased (50%) after treatment; serum IgM and IgA were within normal range and were not affected by treatment High levels of IFN-gamma (5-23 pg/ml) were initially detected in serum, which decreased after treatment (0.1 pg/ml). Furthermore, low levels of IL-4 (0.2 pg/mL) were detected before treatment, which were not detected after treatment (0.1 pg/ml). In contrast, before treatment, IL-12 and TNFalpha were not detected in serum; however after treatment the levels of IL-12 and TNFalpha dramatically increased. Prednisone and azathioprine treatment decreased total serum IgG, IgE, IFN-gamma and IL-4 levels, and blood CD19+ and CD23+ cells; however serum IL-12, TNFalpha and blood CD4+ T cells increased with treatment. Understanding immunomodulation in autoimmune hepatitis will provide better insight and mechanisms of this disease and may tailor more effective therapeutic intervention.

Summary In addition to allergy and parasitic infections, immunoglobulin E (IgE) has been shown recently to possess anti-viral and anti-cancer effects. We investigated serum levels of IgE, its low-affinity receptor, soluble CD23 (sCD23) in patients with pancreatic cancer and the effect of IgE against pancreatic cancer cells. Twelve patients were evaluated for pancreatic cancer by imaging and confirmed by biopsy. Fifteen healthy volunteers served as controls. Serum Igs (IgG, IgM, IgA, IgE) and sCD23 levels were determined (enzyme-linked immunosorbent assay, nephelometry) and the presence of cancer-specific IgE was assessed (fluorescence microscopy, Western blot). IgE anti-cancer activity was determined by antibody-dependent cell-mediated cytotoxicity (ADCC). Serum levels of IgE and sCD23 were elevated significantly in patients with pancreatic cancer versus controls, whereas no differences were observed in other Ig isotypes (IgG, IgM, IgA). Flow cytometry and immunofluorescence microscopy demonstrated similar presence of IgG and IgE pancreatic cancer Igs. However, Western blot analysis indicated differences in IgG and IgE antigen-specific antibodies; IgE antibody recognized a 50 kD protein. ADCC studies demonstrated that serum and purified IgE-mediated cytotoxicity against pancreatic cancer cells, effects which were reversed with anti-IgE neutralizing antibody and IgE depletion (immunoaffinity); greater cytotoxicity was observed in patient serum when compared with healthy controls. These data suggest that IgE and sCD23 may serve as useful biomarkers for patients with pancreatic cancer and may be important in the immune response to this disease in that IgE-directed therapy may help to direct treatment.

Immunoglobulin (Ig) E may provide immunity against Borrelia burgdorferi infection (Lyme disease) in children which lasts throughout adulthood. We investigated the presence and persistence of IgE anti-B. burgdorferi antibodies (Abs) in paediatric patients infected with Lyme disease over time. Serum immunoglobulin levels, presence of IgG and IgE anti-B. burgdorferi components, and distributions of blood T, B and natural killer lymphocyte subsets were studied in B. burgdorferi-infected and -uninfected children (nephelometry, UniCAP Total IgE Fluoroenzymeimmunoassay, Western blot, flow cytometry). Total serum IgM, IgG, IgE and IgA levels, and distributions of blood lymphocytes (CD4(+), CD8(+), CD19(+)) of both groups, excluding CD8(+)CD60(+) T cells, were within normal ranges. However, infected, but not uninfected children made IgG anti-B. burgdorferi proteins p18, p31, p34, p41, p45, but not IgG anti-p60, and IgE anti-B. burgdorferi proteins p31, p34, p41, p45, p60, but not IgE anti-p18. These proteins were also detected in an infected child 1 year post-infection. Interestingly, CD8(+)CD60(+) T-cell numbers were significantly increased (fourfold) in infected, compared with uninfected, patients (P=0.001). These results demonstrate that specific IgE anti-B. burgdorferi Abs are generated and persist in children with Lyme disease and that CD8(+)CD60(+) T cells may play an important role in these responses.

AIM: To investigate the effect of age on severity of acute pancreatitis (AP) using biochemical markers, histology and expression of the protective pancreatitis-associated proteins (PAPs). METHODS: AP was induced via intraductal injection of 4% sodium taurocholate in young and old rats. Sera and pancreata were assayed at 24 h for the parameters listed above; we also employed a novel molecular technique to assess bacterial infiltration using polymerase chain reaction to measure bacterial genomic ribosomal RNA. RESULTS: At 24 h after induction of AP, the pancreata of older animals had less edema (mean ± SE histologic score of young vs old: 3.11 ± 0.16 vs 2.50 ± -0.11, P < 0.05), decreased local inflammatory response (histologic score of stromal infiltrate: 3.11 ± 0.27 vs 2.00 ± 0.17, P < 0.05) and increased bacterial infiltration (174% ± 52% increase from sham vs 377% ± 4%, P < 0.05). A decreased expression of PAP1 and PAP2 was demonstrated by Western blotting analysis and immunohistochemical staining. There were no differences in serum amylase and lipase activity, or tissue myeloperoxidase or monocyte chemotactic protein-1 levels. However, in the most-aged group, serum C-reactive protein levels were higher (young vs old: 0.249 ± 0.04 mg/dL vs 2.45 ± 0.68 mg/dL, P < 0.05). CONCLUSION: In older animals, there is depressed PAP expression related to a blunted inflammatory response in AP which is associated with worsened bacterial infiltration and higher C-reactive protein level; this may explain the more aggressive clinical course.

We previously reported that minocycline treatment of allergic asthmatic patients had oral steroid sparing effects and improved their clinical status and that minocycline suppressed in vitro induction of IgE responses by their PBMC. The effect of minocycline on human or animal IgE responses in vivo has not been studied. Allergic asthmatics (serum IgE: 505 +/- 535 IU ml(-1)) were given minocycline (150 mg po to 250 mg po BID) as add-on therapy to standard care for up to 10 months; control subjects (IgE: 405 +/- 472 IU ml(-1)) received standard care (n = 6 per group). Serum immunoglobulin (IgM, IgG, IgE and IgA) levels were determined monthly (Nephelometry, Unicap Total IgE Fluoroenzyme immunoassay). BALB/c mice (n = 6 per group) were injected intraperitoneally with benzylpenicilloyl(14)-Keyhole limpet hemocyanin (BPO(14)-KLH) in alum on days 0, 21 and 42, fed with minocycline or doxycycline (10-100 mg kg(-1)) on day 44 and numbers of BPO-specific IgG(1), IgE and IgA antibody-forming cell (AFC) in mesenteric LN and spleen and serum immunoglobulin levels were determined on days 46-70 (enzyme-linked immunosorbent spot assay, ELISA). The ability of minocycline or doxycycline to suppress in vitro induction of murine memory IgE responses also was investigated. Minocycline strongly suppressed serum IgE levels of allergic asthmatics (9% per month) (P = 0.012). Minocycline (and doxycycline) also strongly suppressed peak murine IgE AFC and serum IgE responses (>95, approximately 75%, respectively) and in vitro induction of memory IgE responses by murine mesenteric LN and spleen cells (>95%). Tetracycline suppression of all human and murine IgE responses was IgE isotype specific. Suppression of murine IgE responses in vivo was dose dependent and lasted 5-7 days.

IgE levels in cord blood have been investigated as predictors of atopy, but no definitive findings have been made. Other factors, including cells and/or cytokines may serve as predictors of this disease. Cord blood and peripheral blood was obtained at birth and at 7 months of age, respectively, from children (n = 2) with a family history of allergy. Cells in cord blood and peripheral blood were phenotyped and levels of serum immunoglobulins (IgM, IgG, IgA and IgE) were determined. In addition, placentas from these pregnancies were obtained and stained for IgE+ cells and CD8+CD60+ T cells. We found immunoglobulin levels were within normal ranges although IgE levels were negligible in cord blood and at 7 months of age. Similar numbers of CD8+ T cells and CD19+ B cells were detected in cord blood and at 7 months of age. However, CD4+ T cells increased (twofold) and CD16+/CD56+ natural killer precursor cells decreased (twofold) at 7 months of age. CD8+ T cells in their cord blood and at 7 months of age comprised of50% CD8+CD60+ T cells. Cord blood cells expressed epsilon-specific mRNA and mRNA for interleukin-2 (IL-2), IL-4, IL-10 and interferon-gamma (IFN-gamma) but not IL-6. At 7 months of age, peripheral blood mononuclear cells expressed epsilon-specific mRNA and mRNA for all cytokines. In the placental membrane, we detected IgE+ cells, while CD8+CD60+ T cells were detected in the chorionic villi. CD8+CD60+ T cells, cells expressing epsilon-specific and IL-6-specific mRNA may contribute to the pathobiology and provide important prognostic indicators of atopy.

CD8+CD60+ T cells (80–98% CD45RO+; 20% CD23+) are significantly increased in the blood of serum IgE+ ragweed-sensitized (RS) compared with serum IgE-nonatopic humans (p = 0.001). CD8+CD60+ T cells of the RS patients produced IL-2, IL-4, IL-10, IL-12, IFN-α. and IFN-γ, but not IL-6 or IL-13. When their PBMC were cultured with ragweed Ag (RA), peak IgE responses occurred on day 10; none was induced with non-cross-reacting or without Ag; nonatopic PBMC did not respond to any stimulant. When either CD4+ or CD8+CD60+ T cells were depleted from RS PBMC before culture with RA, no IgE responses were induced. If purified CD4+ T cells or low numbers of CD8+CD60+ T cells were added back to the depleted PBMC, IgE responses were restored. However, higher numbers of CD8+CD60+ T cells totally suppressed IgE responses. Total suppression also was obtained when RS PBMC were cultured with RA and either anti-IL-2, IL-4, IL-10, IL-12, IFN-γ (all concentrations), or IFN-α (low concentrations), but not anti-IL-6 or IL-13. Higher concentrations of anti-IFN-α potentiated IgE responses.

S U N D A Y 361 The Association Between Vitamin D Insufficiency and Children with Asthma Ji Tae Choung, MD, Yu-Ri Kim, Sungchul Seo, Young Yoo, MD, PhD; The Environmental Health Center for Asthma; Korea Univ. Medical Center, Seoul, Department of Biochemistry and Molecular Biology, Korea University Medical School & College, Korea, Seoul, The Enviornmental Health Center for Asthma, Korea University, Seoul, Department of Pediatrics, College of Medicine, Korea University, Seoul, South Korea. RATIONALE: Recent epidemiologic studies have shown that the lack of vitamin D levels may be associated with higher asthma prevalence. However, little clinical evaluation was performed in Korea. This study aimed to explore the association of vitamin D levels in serum with asthmatic children in Korea. METHODS: A total of 64 children (34 asthmatic children and 30 healthy children) aged 6-14 years were recruited. Serum 25-hydroxy vitamin D3 (25-OH vitamin D) levels were measured and compared between the both groups. Pulmonary function tests (PFT) as well as questionnaire survey including time of outdoor activity were performed as well. The relationship between serum 25-OH vitamin D levels and the results of PFT as well as time of outdoor activity were examined in asthmatic patients. RESULTS: Serum 25-OH vitamin D levels in asthmatic patients (16.66 4.2 ng/ml) were significantly lower than that in healthy controls (24.2 6 6.8 ng/ml) (P

We have recently found that the tetracycline minocycline suppresses inflammatory responses in serum immunoglobulin (Ig)E-positive asthmatic patients, and that IgE levels can decrease in these patients. The mechanism by which minocycline suppresses these responses is unknown.We have now investigated the ability of the tetracyclines, minocycline and doxycycline, to regulate IgE responses of peripheral blood mononuclear cells (PBMC) obtained from serum IgE-positive asthmatic patients.The distributions of CD3+, CD4+, CD8+, and CD19+ lymphocytes in peripheral blood of serum IgE-positive asthmatic patients and IgE-negative nonasthmatic controls, and cytokine-specific mRNA expression by their PBMC were determined by flow cytometry (reverse transcriptase-polymerase chain reaction). Serum Ig levels also were determined (nephelometry, fluoroenzymeimmunoassay, enzyme-linked immunoadsorbent assay; n = 7/group). PBMC (1.5 x 10(6)/mL) were cultured with anti-CD40 monoclonal antibody and recombinant human interleukin-4 in the presence/absence of minocycline or doxycycline (0.1 to 10 microg/mL), and IgE levels in supernatants determined on days 0, 3, and 10 (enzyme-linked immunoadsorbent assay).Asthmatic and nonasthmatic subjects had similar numbers of blood CD4+ T cells (779/mm3 +/- 73 and 766 +/- 115, respectively) and CD19+ B-cells (239/mm3 +/- 35 and 379 +/- 95, respectively); however, CD8+ T cell numbers were decreased in asthmatic compared with nonasthmatic subjects (378/mm3 +/- 66 and 568 +/- 53, respectively; P = 0.045). High IgE levels were detected in supernatants of asthmatic PBMC on day 10 (28 ng/mL +/- 12), whereas control IgE levels did not change (2.5 ng/mL). When either minocycline or doxycycline was included in culture, IgE production by asthmatic PBMC was strongly suppressed in dose-dependent fashion on day 10 (80% with 10 microg/mL); control IgE did not change (2.5 ng/mL).The results are consistent with the idea that the therapeutic benefits obtained by asthmatic patients from minocycline may, in part, result from IgE suppression.

We previously identified a group of long-term pediatric survivors who had acquired HIV-1 through maternal transmission; had not received antiretroviral therapy; are now8 years old, in good health, and with no opportunistic infections; and have not failed to thrive, although they have greatly decreased numbers of blood CD4+ T cells (500/mm(3)). All the children have elevated total serum IgE levels (210-2475 IU/ml) and make anti-HIV-1 IgE or IgE directed against non-HIV-1 specificities (radioimmunoassay, Western blot assay); they have no detectable antigenemia. We have now studied the ability of anti-HIV-1 IgE in serum obtained from these children to regulate (1) production of HIV-1 by interleukin 2/phytohemagglutinin (IL-2/PHA)-stimulated peripheral blood mononuclear cells (PBMCs) taken from HIV-1-seronegative donors and infected with a T cell-tropic clone of HIV-1, and (2) transmission of a primary HIV-1 strain from adult AIDS patients to uninfected IL-2/PHA-stimulated PBMCs (p24 core antigen production). High levels of HIV-1 production were observed when PBMCs were cultured for 5 days in the presence of HIV-1-seronegative donor serum that was either IgE positive or IgE negative (IgE,100 or100 IU/ml, respectively). HIV-1 production also was observed when PBMCs were cultured with HIV-1-infected donor serum that either contained IgE directed against non-HIV-1 specificities or was IgE negative; these levels were 40% less than those seen with sera from the HIV-1-seronegative donors. Far greater inhibition of virus production was observed if the serum in culture contained anti-HIV-1 IgE (95%). Virus neutralization did not appear to account for the inhibition obtained with anti-HIV-1 IgE-containing serum because virus production was not suppressed in cultures to which serum was added immediately preinfection (10%), but was strongly suppressed when serum was added 1.5 hr postinfection (95%). The inhibition of virus production obtained with serum containing anti-HIV-1 IgE was reversed when (1) serum was depleted of IgE (immunoaffinity), but not when it was depleted of IgG (protein G-Sepharose) before inclusion in culture postinfection, (2) anti-IgE, but not anti-IgG, was included in culture, or (3) serum was heat treated before culture. The results indicate that serum from certain HIV-1-infected pediatric long-term survivors contains agents that inhibit HIV-1 production in vitro, and that these agents include anti-HIV-1 IgE. They suggest that a cytotoxic event, rather than virus neutralization, plays an important role in anti-HIV-1 IgE-mediated inhibition of virus production.

Background: An elevated IgE level and increased production of T H2 cytokines are factors associated with poor prognosis in HIV infection. We report a pediatric long-term survivor of vertically acquired HIV infection with a normal CD4 count and a low viral burden despite the lack of antiretroviral therapy and a phenotype resembling hyper-IgE syndrome. Objective: We sought to characterize the patient's T H1 versus T H2 cytokine profile and anti-HIV–specific immune responses. Methods: Supernatants collected from cultures of peripheral blood T cells stimulated with phorbol myristate acetate plus ionomycin were assayed for T H1 and T H2 cytokines by means of ELISA. Specific IgE antibodies were determined by immunoblot. HIV-specific cytotoxic T-lymphocyte responses were measured from cell lysis by fresh T cells of autologous B-lymphoblastoid cells expressing recombinant HIV proteins. Results: Patient CD4 + T cells secreted significantly more T H2 cytokines, IL-4 ( P P < .03), than HIV-infected and seronegative control cells. No difference was noted in T H1 cytokine production. IgE specific for HIV gp160, p24, p17, and p66 proteins and Aspergillus fumigatus was detected in patient sera. Despite predominance of T H2 cytokines, HIV-specific cytotoxic T-lymphocyte activity was vigorous. Conclusions: The patient demonstrated predominantly T H2 cytokine production in vitro. Unlike other patients with HIV who have hyper-IgE and increased T H2 cytokine production, our patient has maintained HIV-specific immune responses, a low viral load, and a normal CD4 count without antiretroviral therapy. These findings support a diagnosis of primary hyper-IgE syndrome. Presence of anti-HIV–specific IgE may represent a protective mechanism against HIV replication in our patient. (J Allergy Clin Immunol 1999;104:1045-51.)

The ability of interleukin (IL)-6 or interferon-α (IFN-α) to regulate expression of low-affinity Fc receptor (CD23) and serum levels of CD23 was studied in benzylpenicilloyl-keyhole limpet hemocyanin-sensitized BALB/c mice at the peak of a haptenspecific immunoglobulin E (IgE) antibody-forming cell (AFC) response. These responses are analogous to those observed in human atopic disease. To induce peak IgE responses, mice were injected intraperitoneally with BPO-KLH (10 μg) in aluminum hydroxide gel (alum) on days 0, 21, and 42. On day 44, mice were injected subcutaneously with IL-6 (100-1000 U) or IFN-α (1000-10,000 U). On day 46, numbers of CD23+ lymphocytes in Peyer’s patches (PP), mesenteric lymph nodes (MLN), and spleen and levels of soluble CD23 in serum were determined (flow microfluorimetry and enzyme-linked immunosorbent assay, confirmed by competition assay). Data are expressed as percent total cells or as optical density at 490 nm. IFN-α treatment strongly suppressed (up to 100%) numbers of CD23+ cells exclusively in PP (i.e., numbers of CD23+ cells in MLN and spleen were unchanged) whereas serum levels of soluble CD23 were dramatically increased (60%). IL-6 treatment had no effect on either numbers of CD23+ lymphocytes or on serum levels of soluble CD23. The data suggest that the mechanism(s) by which IFN-α, but not IL-6, regulates IgE responses involves suppression of CD23 expression on lymphocytes in PPs and supports a central role for these organs in regulation of IgE responses in vivo.