Intravenous immunoglobulin treatment increased live birth rate in a Spanish cohort of women with recurrent reproductive failure and expanded CD56(+) cells.

PROBLEM: Natural killer (NK, CD3(-)CD56(+)/CD16(+)) and NKT-like cells (CD3(+)CD56(+)/CD16(+)) activity is considered among the key factors for reproductive success. In the absence of immunological screening, beneficial effects of intravenous immunoglobulin (IVIG) in preventing recurrent reproductive failure (RRF) have not been reported. Here, we analyse the IVIG influence on pregnancy success in women with RRF and circulating NK or/and NKT-like cells expansion. METHOD OF STUDY: One hundred fifty-seven women with previous recurrent miscarriage and/or recurrent implantation failure after in vitro fertilization were consecutively studied. Sixty-four patients with CD56(+) cell expansion, no apparent underlying disease and who maintained their desire to conceive were selected. Forty of them received IVIG during pregnancy. RESULTS: Overall, the clinical pregnancy rate for the women under IVIG therapy was 92.5% and the live birth rate was 82.5%. Significantly lower pregnancy and live birth rates (25% and 12.5%, respectively) were observed for the patients with recurrent pregnancy loss and NK/NKT-like cells expansion without IVIG. After three cycles of IVIG, NK cell percentages decreased significantly and these values persisted throughout gestation. CONCLUSION: Intravenous immunoglobulin therapy for women with RRF and NK or NKT-like cell expansion was a safe and beneficial therapeutic strategy that associated with high clinical pregnancy and live birth rates.

Long-term remission of severe refractory dermatopolymyositis with a weekly-scheme of immunoglobulin followed by rituximab therapy.

We report on a 44-year-old woman affected by dermatopolymyositis resistant to conventional therapies who experienced long-term clinical improvement and remission after treatment with intravenous polyvalent immunoglobulin in a weekly schedule followed by rituximab therapy.

Risk factors for developing de novo autoimmune hepatitis associated with anti-glutathione S-transferase T1 antibodies after liver transplantation.

De novo autoimmune hepatitis (de novo AIH) is a rare form of graft dysfunction that develops after liver transplantation (LT) in patients transplanted for conditions other than autoimmune disorders. Although characterized by biochemical, serological, and histological features of AIH, de novo AIH is sometimes associated with atypical serum autoantibodies, many of which are directed against glutathione S-transferase T1 (anti-GSTT1). GSTT1 donor/recipient genotype mismatch has been suggested as a necessary condition for the appearance of autoantibodies and de novo AIH. However, clinically evident disease is not observed in all patients with anti-GSTT1 antibodies. We examined the incidence of de novo AIH and its conditioning (risk) factors in patients with anti-GSTT1 antibodies. Anti-GSTT1 autoantibodies were detected in 29 of 419 [6.9%; 95% confidence interval (CI), 4.9-9.8] consecutive adult LT recipients with donor/recipient GSTT1 mismatch. Twenty of 27 assessable patients (74%) developed de novo AIH after a median follow-up of 26 months (95% CI, 19.2-32.8). The probability of de novo AIH was 11%, 44%, and 60% 12, 24, and 36 months after LT, respectively. No relationship emerged between de novo AIH and recipient gender, donor and recipient age, rejection episodes, immunosuppressive regime, allelic GSTT1 expression, human leukocyte antigen distribution, or cytomegalovirus infection. Multivariate analysis identified male donor [hazard ratio (HR), 3.3; 95% CI, 1.18-9.26; P = 0.018], nonalcoholic etiology (HR, 4.67; 95% CI, 1.64-13.3; P = 0.002), and high anti-GSTT1 titer (HR, 2.98; 95% CI, 1.04-8.57; P = 0.035) as independent predictors of de novo AIH. Most patients with anti-GSTT1 antibodies and donor/recipient GSTT1 mismatch developed clinically evident de novo AIH after LT. The risk of developing the disease was increased by male donor gender, nonalcoholic etiology of original liver disease, and a high anti-GSTT1 titer.

Association between anti-cyclic citrullinated peptide antibodies and ischemic heart disease in patients with rheumatoid arthritis.

OBJECTIVE: Patients with rheumatoid arthritis (RA) have an increased risk of cardiovascular disease that may not always be related to the presence of traditional cardiovascular risk factors. The aim of this study was to determine if anti-cyclic citrullinated peptide (anti-CCP) antibodies are associated with cardiovascular disease in patients with RA. METHODS: Anti-CCP antibodies were determined by enzyme-linked immunosorbent assay in the earliest serum sample available from 937 patients with a diagnosis of RA. We studied the relationship between anti-CCP antibodies with traditional cardiovascular risk factors and cardiovascular events. RESULTS: We found positive anti-CCP antibodies (>25 units/ml) in 672 patients (71.7%). There was no association between the anti-CCP antibodies and cardiovascular risk factors such as smoking, hypertension, dyslipidemia, being overweight, or diabetes mellitus. However, patients who had positive anti-CCP antibodies experienced more frequent ischemic heart disease (6.5% versus 2.6%; odds ratio [OR] 2.58, 95% confidence interval [95% CI] 1.17-5.65) and had higher mortality rates (11.2% versus 6.8%; OR 1.72, 95% CI 1.01-2.91). Similar results were obtained when we considered anti-CCP titers 20-fold higher (>500 units/ml). Multivariable analysis showed that ischemic heart disease is independently associated with positive anti-CCP antibodies (OR 2.8, 95% CI 1.19-6.56; P = 0.009). CONCLUSION: Anti-CCP antibodies in patients with RA are independently associated with the development of ischemic heart disease.

Antibodies against glutathione S-transferase T1 (GSTT1) in patients with GSTT1 null genotype as prognostic marker: long-term follow-up after liver transplantation.

An objective to improve the evolution of transplants is to identify risk biomarkers of morbidity and loss of allograft. In liver transplant (LTX) recipients, an association has been demonstrated between the presence of mismatch for glutathione S-transferase T1 (GSTT1) and the development of de novo immune hepatitis (IH). In 419 LTX patients we analyzed, for a period of 1 to 14 years, the development of "atypical" autoantibodies directed against GSTT1 and their relationship with the mismatch for GSTT1 genotype and with the risk for developing de novo IH. A total of 6.9% LTX recipients had "atypical" autoantibodies and 24 showed mismatch (recipient/donor) for GSTT1 genotype. From this last group, up to 70% developed de novo IH and graft dysfunction after LTX (95% confidence interval: 17.4-37.5 months). In LTX recipients with a GSTT1 null genotype, the evaluation of "atypical" autoantibodies is useful for monitoring the development of de novo IH.

Isolated type 5 antimitochondrial autoantibodies are associated with a history of thrombocytopenia and fetal loss.

OBJECTIVE: To report an unusual case of clinical antiphospholipid syndrome seen with recurrent fetal loss, chronic thrombocytopenia, and recurrent idiopathic thrombocytopenic purpura, in which the only laboratory marker was M5-type antimitochondrial autoantibodies (AMA). DESIGN: Case report. SETTING: University general hospital, tertiary level of clinical care. Institutional practice. PATIENT(S): A 65-year-old woman with antiphospholipid syndrome associated with autoimmune polyglandular syndrome of IIIC type. INTERVENTION(S): Clinical history and biochemical and immunologic markers. MAIN OUTCOME MEASURE(S): The presence of M5-type AMA in clinical antiphospholipid syndrome isolated by indirect immunofluorescence. RESULT(S): During the 10-year follow-up, immunologic studies detected a persistent positive M5-type AMA at high titer (1/640), antithyroid antibodies, anti-gastric parietal cells, and anti-intrinsic factor. Antinuclear, anti-DNA, and antiphospholipid (anticardiolipin, anti-beta(2)-glycoprotein I) autoantibody tests were all repeatedly negative. Results of coagulation studies and negative lupus anticoagulant were normal on several occasions. CONCLUSION(S): Our findings suggest the necessity of determining M5-type AMA in the study panel of antiphospholipid syndrome diagnosis, particularly in the absence of other typical autoantibodies.

Anti-cyclic citrullinated peptide versus anti-Sa antibodies in diagnosis of rheumatoid arthritis in an outpatient clinic for connective tissue disease and spondyloarthritis.

OBJECTIVE: . To compare the diagnostic value of anti-cyclic citrullinated peptide (anti-CCP) and anti-Sa antibodies in serum for prediction of rheumatoid arthritis (RA) in an outpatient clinic for connective tissue diseases and spondyloarthritides. METHODS: A cross-sectional study was carried out to analyze the presence or absence of anti-CCP and anti-Sa antibodies in the sera of 250 randomly selected patients. The disease distribution in the study was as follows: 87 patients had RA (34.8%); 90 (36%) had other connective tissue diseases (CTD); 50 (20%) spondyloarthritis; 19 (7.6%) polymyalgia rheumatica; and 4 (1.6%) juvenile idiopathic arthritis. RESULTS: Anti-CCP antibodies were detected in 63 patients with RA and in 9 patients with other illnesses [sensitivity 72.4%, specificity 94.4%, positive predictive value (PPV) 87.5%]. Anti-Sa antibodies were detected in 38 patients with RA and in 6 patients with other illnesses (sensitivity 43.6%, specificity 96.3%, PPV 86.3%). Anti-CCP and anti-Sa results were discordant in up to 47 of 87 RA patients. No relation between the presence of anti-Sa and higher or lower titers of anti-CCP antibodies was observed. CONCLUSION: The diagnostic value in RA is similar for both antibodies. However, the sensitivity of anti-CCP detection is higher than that of anti-Sa. Our results suggest that presence of anti-Sa antibodies in serum may be useful as a complementary assay when anti-CCP antibodies are negative and RA is suspected.

Pregnancy-induced expansion of regulatory T-lymphocytes may mediate protection to multiple sclerosis activity.

Pregnancy represents a physiological transitory state of immune tolerance to avoid the rejection of the foetus, and concomitantly of stabilisation of many autoimmune diseases, such as multiple sclerosis (MS). Alterations in regulatory T-lymphocytes (T(Reg)) are known to be involved in organ-specific autoimmune disease pathophysiology. Our goal was to quantify CD4+ CD25+ and CD4+ CD25hi+ T(Reg) and activated (CD4+ HLA-DR+ CD38+) T-lymphocytes during pregnancy and puerperium in 13 MS patients in comparison with healthy pregnant and non-pregnant women. During pregnancy, a progressive parallel increase in CD4+ CD25+ T-lymphocytes in healthy pregnants as well as MS pregnant patients was observed. The proportion of T(Reg) was significantly higher in all pregnants than in non-pregnant women (p=0.01), whereas no differences were observed neither in the percentages of total nor activated CD4+ T-lymphocytes. In MS patients, CD4+ CD25+ T-lymphocytes significantly decreased when comparing the third trimester with the puerperal period proportions (p = 0.01), whereas CD4+ CD25hi+ T-lymphocytes significantly increased (p = 0.002). Our findings are consistent with the expansion of circulating regulatory CD4+ CD25+ T-lymphocytes pool with suppressive activity during normal pregnancy and in MS. A different pattern of CD+ CD25hi+ T-lymphocytes between healthy pregnants and MS women, which may represent relevant factors in the activity course of MS.

Short-term sequential analysis of sex hormones and helper T cells type 1 (Th1) and helper T cells type 2 (Th2) cytokines during and after multiple sclerosis relapse.

Multiple sclerosis (MS) is an immune-mediated disease with a clear sex-bias that may be attributed to sex hormones, sex' linked genes or both. Here we sought to determine the evolution pattern of cortisol and sex hormones at MS relapse and 2-months later in 7 male patients with relapsing remitting MS, and whether there was a correlation with a specific Th1 and Th2 cytokine pattern. Our findings indicate the activation of the hypothalamic-pituitary-adrenal axis and the concomitant upregulation of pro- and anti-inflammatory cytokines during relapse. The further increase of sex hormones, in particular estradiol in our male MS patients suggest their possible implication in the physiopathology of the illness and a putative anti-inflammatory and neuroreparatory effect.

[Antiphospholipid antibodies evolving into connective tissue disease: follow-up study of women with recurrent misscarriage]. / Anticuerpos antifosfolipídicos y evolución a conectivopatía: estudio de seguimiento de mujeres con abortos recurrentes.

BACKGROUND AND OBJECTIVE: The aim of this study was to evaluate the clinical and immunologic profile, the rate of progression to connective tissue disease and the possible predictors of evolution in patients with antiphospholipid antibodies and abortions. PATIENTS AND METHOD: In a prospective follow-up study, we determined the prevalence of antiphospholipid antibodies as well as other autoimmune abnormalities and the evolution to connective tissue disease in 200 women with unexplained recurrent abortions. IgG and IgM anticardiolipin antibodies were determined by ELISA and the lupus anticoagulant was determined by means of coagulometric tests. RESULTS: Of 200 women with pregnancy losses, 69 (34.5%) had antiphospholipid antibodies. Thirty-one of 200 women (15.5%) had high or moderate positive anticardiolipin antibodies. During a mean follow-up of 32 months, 9 (13%) antiphospholipid-antibody-positive patients developed features of lupus- like disease or systemic lupus erythematosus. A low total hemolytic complement, increased circulating immune complexes and positive antinuclear antibodies (ANA) were more common in those patients evolving to a connective tissue disorder (p < 0.001, p = 0.003 and p < 0.001, respectively). Positive ANA in women with antiphospholipid antibodies predicted independently the evolution to a connective tissue disorder [Cox proportional hazard model; relative hazard = 4.92, p = 0.04]. CONCLUSIONS: A subgroup of patients with antiphospholipid antibodies and abortions may progress to a connective tissue disorder. A positive antinuclear antibody test result could be useful to identify those patients with antiphospholipid antibodies and abortions who are prone to evolve into a systemic autoimmune disease.

Anticuerpos antifosfolipídicos y evolución a conectivopatía: estudio de seguimiento de mujeres con abortos recurrentes / Antiphospholipid antibodies evolving into connective tissue disease: follow-up study of women with recurrent misscarriage

FUNDAMENTO Y OBJETIVO: El objetivo de este estudio fue la determinación del perfil clínico e inmunológico, la tasa de progresión a conectivopatía y posibles variables predictoras de evolución en pacientes con abortos y anticuerpos antifosfolipídicos. PACIENTES Y MÉTODO: Estudiamos prospectivamente la prevalencia de anticuerpos antifosfolipídicos, otras alteraciones autoinmunitarias y su evolución a conectivopatía en 200 mujeres con aborto recurrente. Los anticuerpos antifosfolipídicos se determinaron mediante técnica de enzimoinmunoanálisis (anticuerpos anticardiolipina IgG e IgM) y por técnicas coagulométricas (anticoagulante lúpico). RESULTADOS: De 200 mujeres con abortos, 69 (34,5 per cent) tuvieron anticuerpos antifosfolipídicos. Treinta y un pacientes (15,5 per cent) tenían anticuerpos anticardiolipina a título moderado-alto. Tras una media de seguimiento de 32 meses, 9 pacientes con anticuerpos antifosfolipídicos (13 per cent) evolucionaron a lupus eritematoso sistémico o a enfermedad lupus like. Se observaron con más frecuencia cifras bajas de complemento hemolítico total, concentraciones elevadas de inmunocomplejos circulantes y anticuerpos antinucleares positivos en las pacientes con anticuerpos antifosfolipídicos que evolucionaron a conectivopatía frente a las que no lo hicieron (p < 0,001, p = 0,003 y p < 0,001, respectivamente). La positividad de anticuerpos antinucleares en las pacientes con anticuerpos antifosfolipídicos predijo de forma independiente la evolución a conectivopatía mediante análisis de regresión de Cox (riesgo relativo = 4,92; p = 0,04).CONCLUSIONES: Un subgrupo de pacientes con abortos y anticuerpos antifosfolipídicos puede evolucionar a conectivopatía. La positividad de los anticuerpos antinucleares podría utilizarse para identificar a pacientes con abortos y anticuerpos antifosfolipídicos positivos que a lo largo de su evolución pueden presentar síntomas indicativos de evolución a conectivopatía sistémica (AU)

Anti-Sa sera from patients with rheumatoid arthritis contain at least 2 different subpopulations of anti-Sa antibodies.

OBJECTIVE: Anti-Sa antibodies have been described to be a highly specific marker for rheumatoid arthritis (RA). We demonstrate the existence of 2 different subsets of anti-Sa antibodies, only one of which is specific for RA. Our objective was to purify the Sa antigen, and to achieve partial characterization of these proteins. METHODS: Saline extract and mitochondrial extract from human placenta were used as antigenic sources. Antigens were purified by immunoaffinity chromatography and studied by ELISA and immunoblotting. RESULTS: Three antigenically active bands of 68, 50, and 46 kDa were purified from the saline extract by immunoaffinity chromatography. Two other bands of 29 and 10 kDa that do not react with anti-Sa antibodies were obtained as well. The 68 kDa band was purified from a mitochondrial extract. These bands are not the same as other known mitochondrial autoantigens such as M2, M4, or M9. The amino terminal sequence of the 68 kDa Sa band is DEPKXEVP. The sequence of the 68 kDa Sa band is not compiled in the databases we searched, as either aminoterminal or internal sequence. Antibodies to 50/46 kDa anti-Sa bands detected by immunoblotting were highly specific for RA, while the 68 kDa antigen reacted in ELISA with sera from patients with RA and systemic lupus erythematosus, the latter showing a marked increase in features of RA. Antibodies directed against the 68 and 50/46 kDa Sa bands fluctuated with time, the 50/46 kDa anti-Sa antibodies present during the active period of the disease, and the 68 kDa anti-Sa antibodies during the remission period. CONCLUSION: At least 2 subsets of autoantibodies are present in anti-Sa sera, one directed against a 68 kDa Sa protein and another to the typical 50/46 bands of the Sa system.

Th1/Th2 cytokine balance and nitric oxide in cerebrospinal fluid and serum from patients with multiple sclerosis.

Tumor necrosis factor (TNF-alpha) and IL-10 are key regulators of the T helper (Th)1/Th2 balance, which is critically skewed in many pathological conditions including immune-mediated inflammatory diseases of central nervous system (CNS) such as multiple sclerosis (MS). Nitric oxide (NO) has been reported to have dual effects on CNS pathology, and to play an important role in MS. We performed a cross-sectional study in 17 randomly selected patients during MS flare-up, and compared levels of TNF-alpha, IL-10 and NO in serum and cerebrospinal fluid (CSF) with the serum values of these mediators in two different control groups, healthy subjects and HIV-infected untreated patients. Serum and CSF values of TNF-alpha, IL-10 and NO were higher in MS patients than in the serum of healthy controls. Two MS patients showed increased levels of NO in CSF, with inversion of the NO(SERUM)/NO(CSF) quotient, which is clearly indicative of an intrathecal production of NO. No correlation among the values of both cytokines and NO, and the laboratory parameters analysed in MS patients (IgG index, presence of IgG oligoclonal bands and albumin quotient) was found. The high levels of TNF-alpha and IL-10 (both in serum and CSF) accompanying an MS attack suggest a simultaneous expression of Th1 and Th2 cytokines as opposed to sequential expression of Th1 followed by Th2 as described in the models of experimental autoimmune encephalomyelitis (EAE). Globally, our results support the inherent heterogeneity of the disease.

Response to steroids in de novo autoimmune hepatitis after liver transplantation.

Graft dysfunction associated with autoimmune phenomena has been recently described in liver transplant recipients without previous autoimmune disease. However, the natural history, diagnostic criteria, and definitive therapeutic approach of de novo autoimmune hepatitis (de novo AIH) are poorly understood. We report 12 cases of de novo AIH 27.9 +/- 24.5 months after liver transplantation: the outcome of 7 patients treated with steroids is compared with a group of 5 nontreated patients. Nontreated patients lost the graft after 5.8 +/- 2.6 months from de novo AIH onset. All treated patients were alive after 48.4 +/- 14 (29-65) months from de novo AIH onset, and none of them lost the graft. However, 5 patients relapsed in relation to steroid tapering. All patients presented an atypical antiliver/kidney cytosolic autoantibody, associated to classical autoantibodies in 10 cases. Histological study showed several degrees of lobular necrosis and inflammatory infiltrate. HLA antigen frequencies and matching were compared with 2 control groups (16 orthotopic liver transplantation [LTX] patients without de novo AIH and 929 healthy blood donors); de novo AIH patients showed a higher prevalence of HLA-DR3 (54.5% vs. 25.9%, P =.04) than healthy controls, which was not observed in LTX patients without de novo AIH. In conclusion, this new disease should be included in the differential diagnosis of unexplained graft dysfunction. In addition, treatment with steroids results in a dramatically improved outcome. However, maintenance therapy is usually required.