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Objectives. To determine the safety and efficacy of a short course of fludarabine combined with cyclophoshamide in lupus nephritis. Methods. A phase I/II open label pilot study. Thirteen patients with active proliferative lupus nephritis received monthly oral boluses of low-dose cyclophoshamide (0.5 gm/m2 on day 1) and subcutaneous fludarabine (30 mg/m2 on days 1–3) for 3–6 cycles. Concomitant prednisone was aggressively tapered from 0.5 mg/kg/day to a low-dose, alternate-day schedule. Patients were followed for at least 24 months after therapy. The primary outcome was the number of patients achieving renal remission defined as stable creatinine, proteinuria Results. The study was terminated early because of bone marrow toxicity. Eleven patients who received at least three cycles were evaluated for efficacy. Ten patients improved markedly with seven patients achieving complete remission and three patients achieving partial remission. There were three serious haematological adverse events during the treatment with one death due to transfusion-associated graft vs host disease. Profound and prolonged CD4 (mean CD4: 98/µl at 7 months and 251/µl at 12 months) and CD20 lymphocytopenia was noted in most patients. Three patients developed Herpes zoster infections. Conclusions. A short course of low-dose fludarabine and cyclophoshamide can induce long-lasting remissions in patients with proliferative lupus nephritis, but severe myelosuppression limits its widespread use. [ABSTRACT FROM AUTHOR]

Objectives: Type I interferon (IFN) is implicated in systemic lupus erythematosus (SLE) pathogenesis. We aimed to identify type I IFN signaling-dependent and -independent molecular pathways in a large population of patients with SLE., Methods: Baseline blood samples from adult patients with moderate to severe SLE from two Phase IIb studies (NCT01438489, n  = 265; NCT01283139, n  = 416) were profiled using whole transcriptome array analyses. Type I IFN gene signature (IFNGS) test status (high or low) was determined using a validated qualitative polymerase chain reaction-based test. IFN-type-specific signatures were developed by stimulating healthy blood with IFN-β, IFN-γ, IFN-λ, IFN-ω, or pooled IFN-α. These, and multiple literature-derived cell type and cytokine pathway signatures, were evaluated in individual and pooled study populations. A Fisher's exact test was used for associations, adjusted for false discovery rate., Results: Whole blood samples from IFNGS test-high patients were enriched versus IFNGS test-low patients for CD40L signaling ( Q  < 0.001), CXC cytokine ( Q  < 0.001), TLR8-mediated monocyte activation ( Q  < 0.001), IgG ( Q  < 0.001), major histocompatibility complex class I ( Q  < 0.001), and plasma cell ( Q  < 0.001) gene expression signatures. IFNGS test-low patients had significant enrichment of eosinophil ( Q  < 0.001), IFN-γ-specific ( Q  = 0.005), and T-cell or B-cell ( Q  < 0.001) signatures. Similar enrichment profiles were demonstrated in patients with primary Sjögren's syndrome, systemic sclerosis, and dermatomyositis., Conclusions: IFNGS test-high patients overexpressed many gene signatures associated with SLE pathogenesis compared with IFNGS test-low patients, reflecting broad immune activation. These results provide new insights into the molecular heterogeneity underlying SLE pathogenesis, highlighting shared mechanisms beyond type I IFN, across several autoimmune diseases., Trial Registration: Clinicaltrials.gov: NCT01438489 and NCT01283139.

A post-hoc analysis of pooled data from two Phase IIb trials (sifalimumab; NCT01283139, anifrolumab; NCT01438489) assessed the clinical significance of a Systemic Lupus Erythematosus (SLE) Responder Index (SRI(4)) response (Week 52) for 736 patients with moderate to severe SLE disease activity (study entry). SRI(4) responders achieved significantly greater improvements in clinical outcome measures (including percentages of patients with a ≥ 7-point reduction in SLE Disease Activity Index (SLEDAI)-2000 (2K), British Isles Lupus Assessment Group "A" or "2B" flare rate, and oral corticosteroid reduction to ≤7.5 mg/day; change from baseline in Physician's Global Assessment; and numbers of SLEDAI-2K organ domains with improvement), as well as in patient-reported outcomes (Patient's Global Assessment, Functional Assessment of Chronic Illness Therapy-Fatigue; Short-Form 36 Health Survey Physical Component Summary, Mental Component Summary, Vitality domain scores) vs. nonresponders. Of patients with abnormal serologies, SRI(4) responders had numerically greater improvements (baseline to Week 52) in anti-double-stranded DNA concentrations vs. nonresponders ( p = 0.051), but there were no differences in C3/C4 concentration changes between the two groups. These results confirm previous findings in a different cohort, indicating that an SRI(4) response is associated with global clinical benefit.

Disorders of human salivary glands resulting from therapeutic radiation treatment for head and neck cancers or from the autoimmune disease Sjögren syndrome (SS) frequently result in the reduction or complete loss of saliva secretion. Such irreversible dysfunction of the salivary glands is due to the impairment of acinar cells, the major glandular cells of protein, salt secretion, and fluid movement. Availability of primary epithelial cells from human salivary gland tissue is critical for studying the underlying mechanisms of these irreversible disorders. We applied 2 culture system techniques on human minor salivary gland epithelial cells (phmSG) and optimized the growth conditions to achieve the maintenance of phmSG in an acinar-like phenotype. These phmSG cells exhibited progenitor cell markers (keratin 5 and nanog) as well as acinar-specific markers-namely, α-amylase, cystatin C, TMEM16A, and NKCC1. Importantly, with an increase of the calcium concentration in the growth medium, these phmSG cells were further promoted to acinar-like cells in vitro, as indicated by an increase in AQP5 expression. In addition, these phmSG cells also demonstrated functional calcium mobilization, formation of epithelial monolayer with high transepithelial electrical resistance (TER), and polarized secretion of α-amylase secretion after β-adrenergic receptor stimulation. Taken together, suitable growth conditions have been established to isolate and support culture of acinar-like cells from the human salivary gland. These primary epithelial cells can be useful for study of molecular mechanisms involved in regulating the function of acinar cells and in the loss of salivary gland function in patients., (© International & American Associations for Dental Research 2014.)

A 38-year-old female patient with systemic lupus erythematosus presented with pulmonary infiltrates and hypoxemia for several months following immunodepleting autologous hematopoietic stem cell transplantation. She was treated for influenza, which was isolated repeatedly from oropharynx and bronchoalveolar lavage (BAL) fluids, and later empirically for lupus pneumonitis, but died 6 months after transplant. Autopsy findings failed to show influenza in the lungs or lupus pneumonitis. A novel generic polymerase chain reaction (PCR)-based assay using degenerate primers identified human coronavirus (CoV) HKU1 RNA in BAL fluid at autopsy. CoV was confirmed by virus-specific PCRs of lung tissue at autopsy. Electron microscopy showed viral particles consistent with CoV HKU1 in lung tissue both at autopsy and from a previous biopsy. Although human CoV HKU1 infection is not usually severe, in highly immunocompromised patients, it can be associated with fatal pneumonia., (© 2011 John Wiley & Sons A/S.)

Sjögren's syndrome is an autoimmune exocrinopathy, mainly affecting the lacrimal and salivary glands, and resulting in ocular and oral dryness (keratoconjunctivitis sicca and xerostomia). The aetiology and pathogenesis are largely unknown, and only palliative treatment is currently available. Data obtained from experimental animal and human studies using biological agents or gene therapeutics can offer insight into the disease process of Sjögren's syndrome. This article reviews the current literature on these approaches and assesses the lessons learnt about the pathogenesis of Sjögren's syndrome.

Objectives: Increased levels of hypomethylated CpG-containing DNA in sera from patients with systemic lupus erythematosus (SLE) may contribute to the initiation and/or perpetuation of the disease. This study characterizes the in vitro response of peripheral blood mononuclear cells (PBMC) from SLE patients to CpG DNA., Methods: Secretion of cytokines and IgM, cell proliferation and up-regulation of co-stimulatory molecules were evaluated in PBMC from SLE patients (n=24) and normal controls (n=24) after stimulation with synthetic oligodeoxynucleotides (ODN) containing CpG motifs., Results: Up-regulation of co-stimulatory molecules and the secretion of interferon-alpha and interleukin-6 (IL-6) in response to CpG ODN was significantly reduced in monocytes and dendritic cells from SLE patients. Secretion of interferon-gamma by natural killer (NK) cells was also reduced. In contrast, the IgM and IL-10 response of B cells to CpG ODN was normal., Conclusion: Monocytes, dendritic cells and NK cells from SLE patients respond abnormally to CpG ODN stimulation, which may contribute to the cytokine imbalance observed in SLE.

Endometriosis is one of the most frequent gynecological diseases. In its development take a part genetical, mechanical and immunological factors. Main symptoms are cycle-dependent pelvic pain, infertility and characteristic morphological alterations ("pelvic mass"). For the diagnosis the laparoscopy plays the leading role. Therapy is based either on the hormonal inhibition of ovarian function (danazol, GnRH agonists), or surgical interventions (operative laparoscopy), as well as the combination of both methods.

129 sera with known antibody titres against E. coli 026 and E. coli 055 strains were tested with the Abbot second generation anti-HTLV III recombinant screening assay. No difference in the O.D. values was found between sera with high, normal and low anti-E. coli titres. In addition, no false-positive reactions were observed with the anti-HIV negative sera containing E. coli antibodies in high titres in a Western blot assay in which recombinant env antigen was applied. These results suggest that E. coli assays in which E. coli-produced recombinant antigens are used.

Polyamine oxidase activity was measured in different compartments of the feto-maternal unit by a radiochemical method. The activity in the retroplacental serum (mainly of intervillous origin) was 20 to 30 times higher than in maternal uterine or peripheral venous blood sera. No activity was found in the fetal cord blood sera. The enzyme level in the maternal peripheral sera fell to undetectable levels within 72 hours post partum. Preliminary data indicate that the enzyme might be produced by the decidua rather than by the placenta. It is suggested that the enzyme may constitute an important part of a supposed local immunological barrier formed at the placental bed to help to protect the fetal allograft from maternal immune rejection through a local suppressive effect on maternal cellular immunity.

Serum samples of 589 haemodialysis patients were screened for HIV antibody by ELISA methods. Of these, 36 samples were found to be repeatedly reactive. None of the 36, however, could be confirmed by competitive enzyme immunoassays and Western blot; therefore, they were considered to be false positive. The sera could be divided in two groups. The sera of Group 1 were designated as the usual type of false positivity, caused most probably by anti-lymphocyte antibodies. In 19 sera, however, a special type of false positivity was found. These sera reacted strongly with the plates coated with the supernatants of HIV-infected cells but not with those of uninfected H9 cells. Three and two sera showed, respectively, positive immunofluorescence reaction with the HIV-infected, but not with the uninfected, H9 and CEM cells. Reactivity to HIV-infected H9 cells could be adsorbed from a part of these samples with lesser amounts of HIV-infected than uninfected H9 cells. This special type of false positivity was observed frequently (7/65) in patients who rejected a kidney graft. These findings suggest that this type of anti-HIV false positivity is due to antibodies reacting with cellular antigens present in HIV-infected but not in uninfected lymphocytes. Their appearance seems to be associated with the immunological activation occurring at graft rejection.

Serum polyamine oxidase activity was determined in 38 women admitted to the Maternity Department of Szombathely, Hungary with clinical signs of threatened abortion between 11 and 22 weeks gestation and who aborted spontaneously with 48 h of admission. Serum polyamine oxidase activity was found to be significantly lower in these women compared with that in women having normal pregnancies of the same gestational age. The higher polyamine oxidase levels which were found in five women who aborted during oxytocin infusion probably resulted from increased contractions expelling enzyme-rich blood from the intervillous space into the maternal circulation. Present evidence is insufficient to explain the full clinical significance of these findings.

The occurrence of first cousin couples was 2.90/00 in Hungary involving three different population samples (about 2,000, 1,000 and 2,000 women after delivery in the capital, two cities, and a city with agricultural characteristics, respectively).

In order to clarify whether HIV-1 core and env antigens are destroyed during pepsin treatment, used previously for detecting HIV-1 core and env antibodies hidden in circulating immune complexes, purified recombinant env and core antigen preparations were treated with pepsin. Core antigen was found to be extremely sensitive to this enzyme. By contrast, the antigenicity of the purified env antigen was not destroyed and was even increased after pepsin treatment, performed under identical conditions. These findings suggest that after pepsin digestion the core-anti-core immune complexes do not reconstitute because of the loss of antigenicity of the core antigen. By contrast, the lack of binding after neutralization to the env antigen of the F(ab')2 fragment of the anti-env antibody, cleaved by pepsin from the immune complexes, is probably due to other factors.

Human retroplacental blood serum significantly (p less than 0.01) suppresses the in-vitro uptake of 3H-thymidine--that is, synthesis of deoxyribonucleic acid--by spontaneously growing human lymphocytes in the presence of exogenous spermine, but only in concentrations with a higher polyamine oxidase activity than that found in maternal peripheral blood serum during pregnancy. These findings together with observations that the placenta is rich in spermine and that interaction of polyamine oxidase and substrate arrests cell proliferation suggest that such interaction might represent a localised immunoregulatory mechanism in the placental bed, which might contribute to the protection of the fetoplacental unit from possible maternal immune rejection.

Polyamine oxidase activity, already shown to be present in maternal serum, was measured in amniotic fluid between 15 and 40 weeks of gestation in 38 samples. The enzyme activity increased as pregnancy progressed, the levels being lower than those found in the maternal serum. Homogenates of chorion, amnion, separately and together, also showed the presence of enzyme activity, the levels being lower than those previously found in the decidua, but substantially higher than those found in the placenta. Ii is suggested that the enzyme found in amniotic fluid might be the consequence of a diffusion from the decidua through the membranes.

The existence of a spermine oxidizing enzyme was established in human pregnancy serum by a radiochemical method, and a cross-sectional study of its quantitative profile between 10 and 40 weeks gestation was undertaken. Though there was a wide scatter of values, the rise in the average activity was continuous and highly significant. The flavoprotein inhibitor, quinacrin, inhibited the enzyme activity, while the pyridoxal phosphate inhibitor, isoniazid, had no significant inhibitory effect suggesting that the enzyme is of tissue origin. The possible role of polyamines and their oxidation in physiological immunoregulation is discussed.

The authors report on the results of Mycoplasma hominis and Ureaplasma urealyticum cultivations performed within two and a half years in 1,421 pregnant women and gynaecological patients. Considering the obstetrical-gynaecological patients Mycoplasma hominis was identified in 7.25% and Ureaplasma urealyticum in 38.11% of cases. No significant difference was found in the incidence of Mycoplasma infection when comparing the results of cultivations of samples obtained from a clinically non-inflamed vagina or cervix with the results of cultivations of samples obtained from the female lower genital tract in the course of an inflammatory disease. In cases of endometritis, salpingitis or cophoritis Mycoplasma hominis could be isolated from the cervix significantly more frequently. The incidence of Mycoplasma hominis infection was also significantly higher in the examined 315 sterile women. The incidence of Mycoplasma hominis and Ureaplasma urealyticum infection did not differ when comparing the 267 symptom-free pregnant women to the other patients. The presence of the pathogenic organisms did not aggravate the unfavourable course of pregnancy. When examining the effectivity of Doxycycline capsule the drug proved to be effective in 78 patients for the treatment of genital Mycoplasma infections.