HLA analysis in patients with degenerative diseases of the temporomandibular joint.

The aim of this study was to determine the presence of HLA alleles, specifically HLA-DR alleles, and to correlate them with clinical and radiological features of patients with degenerative processes (DP) of the temporomandibular joint (TMJ). The final goal was to determine which allele can be used to identify patients having more aggressive forms of the articular pathologies. Thirty-two (32) Caucasian patients with DP of the TMJ were included in the study. The SSOP (Luminex Corp., Austin, TX) method was used to determine class II HLA alleles. The presence of HLA-II DR in patients with DP of the TMJ was 98%. The presence of HLA was significantly higher in patients with DP of the TMJ than in healthy subjects (66%) (p=0.003). HLA DR52 was significantly more frequent in patients than in healthy individuals (40.62% vs. 13.79%, p = 0.041). While the percentage of DR11 positive individuals was also higher among patients than among healthy control subjects, the association with DP of the TMJ was not significant (p=0.220). Patients having the DR52 allele had higher deformation or DP. It was concluded that HLA-DR54 and DR11 alleles are associated with a higher susceptibility to DP of the TMJ, and HLA-DR54 and DR52 are associated with a higher severity of DP.

Direct vesicular transport of MHC class II molecules from lysosomal structures to the cell surface.

Newly synthesized MHC class II molecules are sorted to lysosomal structures where peptide loading can occur. Beyond this point in biosynthesis, no MHC class II molecules have been detected at locations other than the cell surface. We studied this step in intracellular transport by visualizing MHC class II molecules in living cells. For this purpose we stably expressed a modified HLA-DR1 beta chain with the Green Fluorescent Protein (GFP) coupled to its cytoplasmic tail (beta-GFP) in class II-expressing Mel JuSo cells. This modification of the class II beta chain does not affect assembly, intracellular distribution, and peptide loading of the MHC class II complex. Transport of the class II/ beta-GFP chimera was studied in living cells at 37 degrees C. We visualize rapid movement of acidic class II/beta-GFP containing vesicles from lysosomal compartments to the plasma membrane and show that fusion of these vesicles with the plasma membrane occurs. Furthermore, we show that this transport route does not intersect the earlier endosomal pathway.

Estimated risk for developing autoimmune Addison's disease in patients with adrenal cortex autoantibodies.

CONTEXT: Patients with adrenal cortex autoantibodies (ACA) without overt autoimmune Addison's disease (AAD) are at risk of adrenal failure. DESIGN: To assess the contribution of different clinical, immunological, genetic, and functional factors in the progression to AAD, we followed up 100 ACA-positive and 63 ACA-negative patients without AAD for a maximum of 21 yr (mean 6.0 yr, median 4.8). ACA were measured by immunofluorescence and 21-OH autoantibodies (Abs) by RIA. Adrenal function was assessed by measuring basal levels of cortisol, aldosterone, ACTH, renin activity, and cortisol response to ACTH. The risk of developing AAD was calculated using survival and multivariate analyses. RESULTS: AAD developed in 31 ACA-positive patients and one ACA-negative patient. The cumulative risk of disease in ACA-positive patients was 48.5% [95% confidence interval (CI) 40.8-56.1]. The cumulative risk was higher in children than adults (100 vs. 31.9%; P < 0.0001), males than females (68.6 vs. 42.7%; P = 0.006), patients with subclinical rather than normal adrenal function at entry (87.4 vs. 30.1%; P < 0.0001), patients with hypoparathyroidism and/or candidiasis than patients with other autoimmune or nonautoimmune diseases (100 vs. 29.7%; P < 0.0001), and patients with high rather than low-medium ACA titers (62.8 vs. 41.2%; P = 0.12). The presence of human leukocyte antigen (HLA)-DRB1 did not appear to contribute to the prediction of AAD. Adjusted hazard ratios by Cox model for the development of AAD were 3.37 for males (CI 1.38-8.24), 5.23 for hypoparathyroidism and/or candidiasis (CI 1.53-17.92), 3.33 for high antibody titers (CI 1.43-7.78), and 6.15 for impaired adrenal function at entry (CI 2.79-13.57). CONCLUSIONS: These results were used to construct a risk algorithm for estimating the probability of developing AAD from the combination of gender, age, adrenal function, antibody titer, and associated autoimmune disorders at entry. The values of estimated risk could be used to decide appropriate follow-up intervals and future immunointervention strategies.

Isolated precocious pubarche: an approach.

Precocious pubarche (PP) is most often a benign condition secondary to the early appearance of adrenarche. However, PP may be a manifestation of nonclassical adrenal hyperplasia. The incidence of nonclassical adrenal hyperplasia in patients with PP ranges from about 0-40% of cases. Controversy exists as to whether all children with PP should undergo an ACTH stimulation test. The aim of this study was 1) to determine the frequency of mild adrenal enzyme defects in a very large and ethnically homogeneous group of children with isolated PP (typical pubarche); 2) to determine whether clinical data, in particular bone age, and basal hormonal values can help to distinguish patients who are at risk for having adrenal enzymatic defects and thus should have an ACTH test; and 3) to determine which patients diagnosed as having a mild adrenal enzyme defect might require treatment. We studied 171 subjects (135 girls and 36 boys), aged 7 +/- 1.2 (SD) yr, with isolated PP. Thirty-eight normal subjects (18 age-matched and 20 pubertal) were studied as controls. An ACTH stimulation test (Synacthen, 0.25-mg iv bolus) was performed. Blood samples were drawn at baseline and 1 h postinjection. 17 alpha-Hydroxyprogesterone (17OHP), 17 alpha-hydroxypregnenolone (17PGN), dehydroepiandrosterone, androstenedione, testosterone, 11-deoxycortisol, and cortisol were evaluated. Haplotype (HLA) typing was performed in the patients who were diagnosed with nonclassical 21-hydroxylase deficiency (NC21OHD). Using published nomogram standards for the serum 17OHP response to ACTH, 10 patients (5.8%) were diagnosed as having NC21OHD. Seven of 112 patients (6.2%) were diagnosed as having nonclassical 3 beta-hydroxysteroid dehydrogenase deficiency (NC3HSD) on the basis of the following three criteria: stimulated 17PGN levels and stimulated 17PGN/17OHP and 17PGN/cortisol ratios higher than 2 SD above the mean for pubertal controls. None of the patients had stimulated 11-deoxycortisol values greater than 2 SD above the mean of pubertal controls. Nineteen patients (11%) had a stimulated 17OHP response characteristic of the heterozygotes for 21-hydroxylase deficiency. One hundred and thirty-five of 171 patients with no biochemical evidence of an adrenal biosynthetic defect were diagnosed as having precocious adrenarche. Bone age was advanced (> 2 SD for chronological age) in 80% of the patients with NC21OHD, in 71.4% of the patients with NC3HSD, in 58% of the patients classified as heterozygotes, and in 32.6% of the patients with precocious adrenarche. Basal hormone levels were helpful in detecting NC21OHD, but not NC3HSD. All patients with NC21OHD and only 1 with NC3HSD underwent glucocorticoid suppression treatment.(ABSTRACT TRUNCATED AT 400 WORDS)

High prevalence of manifestations of gastric autoimmunity in parietal cell antibody-positive type 1 (insulin-dependent) diabetic patients. The Belgian Diabetes Registry.

Previous studies have shown a high prevalence of gastric parietal cell antibodies (PCA) in type 1 diabetes, which can be accompanied by (sub)clinical autoimmune gastric disease. This study aimed to determine the grade of associated autoimmunity and to assess the pattern of prevalence of PCA by gender, age, duration of disease, age at onset of diabetes, and human leukocyte antigen (HLA) type in an adult type 1 diabetic population. Furthermore, to examine the clinical significance of being PCA positive, manifestations of gastric autoimmune disease were studied in PCA-positive and PCA-negative patients. The population studied consisted of 497 type 1 diabetics (men/women, 252/245; mean age, 40.8 +/- 12.1 yr; mean duration of disease, 16.4 +/- 10.4 yr; mean age at onset, 26.9 +/- 13.5 yr; mean hemoglobin A1c, 8.1 +/- 1.6%). Associated autoantibodies were present in 39% and PCA were present in 20.9% of the subjects, particularly in older patients. Gender, duration, and age at onset of diabetes did not influence the appearance of PCA. Antithyroid peroxidase antibodies (aTPO) were more frequent in PCA-positive patients than in those without PCA (33.6% vs. 22.4%; P = 0.025), suggesting an association between gastric and thyroid autoimmunity. We could demonstrate an association between PCA and the HLA DR5 haplotype (P = 0.001) as well, but not with HLA DR3 and/or DR4. In the PCA-positive group, iron deficiency anemia was detected in 15.4%, and pernicious anemia was found in 10.5% of subjects. These autoimmune gastric manifestations were significantly more prevalent in PCA-positive diabetics than in PCA-negative subjects, in whom the percentages were 6.9% and 0.5%, respectively (P = 0.01 and P < 0.0001). PCA were prevalent in 84.6% of patients with pernicious anemia. A gastroscopic and anatomopathological examination performed in a subgroup of 30 patients with gastric symptoms revealed atrophic gastritis in 13 of 14 PCA-positive patients and in 9 of 16 PCA-negative subjects (P = 0.04). PCA were present in 59.1% of patients with atrophic gastritis. In conclusion, a high prevalence of parietal cell antibodies and associated autoimmune gastric disease is present in PCA-positive type 1 diabetics, recommending its screening. Early detection of PCA and iron deficiency anemia, pernicious anemia, and atrophic gastritis and the subsequent care could reduce the morbidity of type 1 diabetes.

A novel method for generating stable high-level transfectants involving direct immunomagnetic selection for cell-surface epitopes: expression of HLA class-II genes in HeLa cells.

A versatile method that allows efficient detection and selection of both transient and stable transfectants expressing exogenous cell-surface molecules is described and used to generate stable HeLa transfectants expressing each of the human HLA class-II isotypes, specifically the DR1, DQw8 and DPw2 heterodimers. The method combines use of the strong mammalian expression vector, CDM8, and a highly efficient transfection protocol with the powerful technique of immunomagnetic selection. It offers significant advantages in comparison to standard procedures involving co-selection with drug-resistance markers. The transfection efficiency can be assessed 60 h after transfection rather than after three weeks of drug selection. Repeated rounds of immunomagnetic selection applied over the subsequent ten days result in homogeneous populations which express the surface marker of interest stably at high levels, making further subcloning or fluorescence-activated cell sorting unnecessary. Any number of surface products can be transfected into the same cell, the only limitation being the availability of specific monoclonal antibodies (a DP/DR double transfectant is described expressing four exogenous gene products simultaneously). The high sensitivity of immunomagnetic selection and its applicability to large samples allows rescue of transfectants present at very low frequencies. Finally, the technique can be used as a coselection procedure, by analogy with drug coselection, to achieve expression even of non-cell surface products.

Human leukocyte antigen and season of birth in Japanese patients with schizophrenia.

OBJECTIVE: Five Japanese studies, to the authors' knowledge, without exception, have consistently shown an increased frequency of human leukocyte antigen (HLA)-DR1 in patients with schizophrenia. This suggests an association between HLA-DR1 and schizophrenia in the Japanese population. The mechanism of the association is unknown; however, prenatal infections may be involved. The present study explored factors, including winter birth, that might correlate with this mechanism. Age at onset and gender were also studied. METHOD: Factors were compared between Japanese patients with schizophrenia with and in those without HLA-DR1 (N=60 and N=307, respectively). RESULTS: A significantly higher incidence of births in February and March was observed in patients with (31.7%) than those without (15. 6%) HLA-DR1. No association was found between the presence of HLA-DR1 and other variables. CONCLUSIONS: Although this result is preliminary, it may suggest an interaction between HLA and winter birth in the development of schizophrenia in the Japanese population.

HLA antigen frequencies in renal transplant recipients and non-immunosuppressed patients with non-melanoma skin cancer.

An increased frequency of human leukocyte antigen (HLA)-DR1 was found in 49 non-immunosuppressed patients with non-melanoma skin cancer (NMSC), being highest in patients under the age of 60 with multiple squamous cell carcinomas (SCC). Of 266 patients receiving long-term immunosuppression following renal transplantation 46 (17%) were found to have NMSC. No increase in HLA-DR1 was found in renal transplant recipients (RTR) with non-melanoma skin cancer (RTR+C) when compared with matched renal transplant recipients without skin cancer (matched RTR-C), or when compared with healthy controls. There was an increased frequency of DQw2 in RTR+C, most pronounced in RTR with SCC (61.9% compared with 18.75% in matched RTR-C), giving a relative risk of 13.98. We found statistically significant differences in the frequency of a number of HLA antigens on comparing RTR+C with healthy controls, but none of these differences were found when we compared RTR+C against matched RTR-C.

T cell responses to D-penicillamine in drug-induced myasthenia gravis: recognition of modified DR1:peptide complexes.

The anti-rheumatoid drug D-penicillamine (D-pen) has a reactive sulfhydryl group capable of modifying self antigens, and can provoke typical autoantibody-mediated myasthenia gravis (MG), especially in DR1+ individuals. We have selected T cell clones from one such patient that were highly specific for D-pen but not its L-isomer or D-cysteine. Moreover, they were restricted to HLA-DR1, had a Th1 phenotype and used TCR V alpha4.1, V beta6.1. They responded well to blood mononuclear cells prepulsed with D-pen either in the absence of serum or after chloroquine treatment, but not to autologous D-pen-pulsed B cell lines. Thus, D-pen may directly couple to distinctive peptides resident in surface DR1 molecules on circulating macrophages or dendritic cells.

HLA-DR1 is associated with vitiligo.

In Eastern Hungary, vitiligo is found to be associated with HLA-DR1. When other autoimmune disorders are also present, DR3 is also increased.

HLA-DPB1 alleles in patients with rheumatoid arthritis.

The distribution of HLA-DPB1 alleles was studied in 94 adult Caucasian RA patients (65 = seropositive, 29 = seronegative) and 40 normal controls. The DPB1 alleles were defined by oligonucleotide typing of polymerase chain (PCR)-amplified genomic DNA. The prevalence of the DPB1*0402 allele was higher in seropositive RA patients and DPB1*0201 was higher in seronegative RA patients. These differences were not significant, however. It is likely that DPB1 alleles do not play an important role in susceptibility to RA.

Genetic markers in early diabetic retinopathy of adolescents with type I diabetes.

The aim of this study was to evaluate the role of HLA (human leucocyte antigen) class I (A, B, C) and class II (DR) alleles and familial insulin-dependent diabetes mellitus as possible risk markers for early retinopathy in a population of 103 Finnish adolescents with type I diabetes mellitus for 3.6-16.2 years. Fifty-one of the patients (49.5%) had signs of retinopathy in fundus photographs. HLA DR1 was found in 31% of the subjects with retinopathy, but in only 5% of those without retinopathy (p = 0.02). The corresponding figures for HLA DR1/4 were 17% and 2.6%, respectively (p = 0.22). The frequency of HLA DR3, DR4, or DR3/4 heterozygosity did not differ between the two groups of patients. Signs of early retinopathy showed thus an association with the presence of the HLA DR1 allele, and a mild protective effect of the HLA A9 and B40 alleles was indicated. Other HLA A, B, C, or DR alleles did not have any effect on the risk for early development of retinopathy, neither had a positive family history of type I diabetes.

HLA antigen frequencies in end-stage idiopathic and ischaemic cardiomyopathy.

We investigated the distribution of HLA antigens among 413 patients with ischaemic heart disease or dilated cardiomyopathy referred for cardiac transplantation to determine if possession of certain HLA antigens predisposed to end-stage heart failure. Of the patients studied, 234 had ischaemic heart disease (218 males), mean age 49 years (SD 7.1) and 179 patients had dilated cardiomyopathy (150 males), mean age 39 years (12.8). The control group comprised 2041 kidney donors reported to the United Kingdom Transplant Service between July and August 1985. We found a significant excess of HLA DR1 (odds ratio 1.64, 95% CI 1.16-2.33, attributable risk 5.0%) and DR5 antigens (odds ratio 1.47, 95% CI 0.99-2.18, attributable risk) among patients with dilated cardiomyopathy but not of HLA DR4 as previously reported. We found a lower frequency than expected of HLA B21 (10.44 expected, none observed) among patients with ischaemic heart disease but no other significant differences. This study provides some support for the concept of the risk of developing end-stage heart failure due to dilated cardiomyopathy being associated with possession of HLA DR1 and DR5, but no such evidence in ischaemic heart disease. Larger multi-centre studies are required to confirm the validity of these findings.

Histocompatibility antigen studies in women with recurrent miscarriages and Müllerian uterine anomalies.

OBJECTIVE: To determine whether the HLA alleles DR1, DR3, DR4 and DR10 which have been suggested to be risk markers for unexplained recurrent miscarriages also play a part in women with recurrent miscarriages with Müllerian uterine fusion anomalies. STUDY DESIGN: HLA-DR typing was undertaken in 28 women with recurrent miscarriage who had been surgically treated for Müllerian anomalies and in 360 controls. In the study group, outcome of pregnancies after surgery was correlated to the results of the HLA typing. RESULTS: In the study group, 61% were positive for HLA-DR1, -DR3, -DR4 or -DR10 compared with 65% of the controls (not significantly different). Among patients positive for these HLA risk markers, 64% of the pregnancies after surgery miscarried compared with 13% in those negative (P<0.005, RR for miscarriage=4.8, 95% CI= 1.3-18.0). CONCLUSION: The proposed risk HLA markers for unexplained recurrent miscarriage also seem to display a negative impact on pregnancy outcome in patients with recurrent miscarriages with Müllerian uterine anomalies.

Lymphocyte response in vitro to Plasmodium falciparum merozoite antigens in donors from a holoendemic area.

Crude merozoite antigens from P. falciparum were used to analyse the proliferative response of peripheral blood mononuclear cells from 114 inhabitants of the village of Dielmo (Senegal, West Africa), who are exposed continuously to malaria transmission. The high or low responses to merozoite antigens obtained in lymphocyte stimulation assays were correlated to the presence or absence of parasites, to the IFN-gamma production and to the HLA-phenotype. High responders produced high levels of IFN-gamma while low responders did not secrete IFN-gamma (23/27). The two HLA phenotypes HLA-B51 and HLA-DR1 were significantly associated with high response (p < 0.05).

[Sarcoidosis synchronously detected in identical twins].

We report on the synchronous detection of sarcoidosis in identical 22-year-old male twins. The patients visited a nearby physician because of fever and cough, were found to have abnormal chest shadows and thus admitted to our hospital. Both had swollen cervical and hilar nodes and lesions in both left and right lung fields, and also presented similar clinical symptoms. However, bilateral uveitis was observed only in the elder twin. Because epithelioid cell granulomas without caseous necrosis were histopathologically observed in tissue specimens from their cervical lymph nodes, the patients were given a diagnosis of sarcoidosis. Although they were positive for the HLA-D antigens DR 2 and DR 12, they were negative for DRw 52. an antigen that is considered to be quite common in Japanese sarcoidosis patients. Genetic factors were thought to be involved in the onset of sarcoidosis in these twins.

[Comparison of HLA class I and DR class II antigen frequency in Polish patients, in relation to different stages of the disease]. / Porównanie czestosci wystepowania swoistosci serologicznych antygenów klasy I i grup alleli DR klasy II ukladu HLA, wsród chorych na sarkoidoze w populacji polskiej, w odniesieniu do stadium choroby.

The aim of this study was to analyze the association between HLA class I and class II DR frequency in the different stages of sarcoidosis in Polish population. 88 patients and 30 healthy controls have been typed. Patients were divided into three groups depending on radiological findings. In the first group were 28 cases presenting the regression of the disease. In the second were 33 patients in stable stage II or III and in the third group 27 patients with pulmonary fibrosis (stage IV). The typing was performed by NIH method using commercial sera. There were no statistically significant differences between studied group in HLA-A class I. The frequency of HLA B-18 was statistically more frequent in patients with sarcoidosis compared to healthy controls. HLA-DR1 was not present in third group of patients and the difference was significant compared to healthy controls.

Low and stable prevalence of rheumatoid arthritis in northern France.

OBJECTIVES: Few data are available on the prevalence of rheumatoid arthritis (RA) in France. Results of the Epidémiologie des rhumatismes inflammatoires (EPIRHUM-2) study suggest a lower prevalence in northern France (0.13%) than nationwide (0.31%) or in southern France (0.66%). Here, our objective was to confirm the lower prevalence of RA in northern France than in the rest of the country or in Europe. METHODS: We used the universal health insurance database to identify patients with RA in northern France (Nord-Pas de Calais region) in 2005. RESULTS: Seven thousand one hundred and twenty-eight patients of the 3,617,224 individuals receiving health insurance under the plan for salaried workers in the Nord-Pas de Calais region (89.3% of the population in the region) were listed as receiving free care for RA, yielding a prevalence of 197.1/100,000 population (female-to-male ratio, 2.99:1; mean age, 60.8 years). DISCUSSION: The prevalence of RA in northern France (0.197% in 2005) is lower than in southern France and northern Europe; thus, the prevalence gradient in France is in the opposite direction to the decreasing north-to-south gradient previously described in Europe. Although environmental and genetic factors involved in the pathogenesis of RA may be involved, the main explanation to the low prevalence in northern France may be the young age of the population.

Limited heterogeneity of the HLA class II contribution to susceptibility to rheumatoid arthritis is suggested by positive associations with HLA-DR4, DR1 and DRw10.

Most patients with rheumatoid arthritis (RA) exhibit an HLA class II epitope found on most DR4 and DR1 molecules. We have investigated the possibility of other associations being present in the minority of patients lacking these antigens. One hundred and eighty patients with classical or definite RA and 100 controls were assigned HLA-DR, DQ and DP types by a DNA-based system using sequence-specific oligonucleotides to probe amplified class II alleles amplified by the polymerase chain reaction. The expected associations with DR4 (relative risk 10, P less than 0.0001) and DR1 (relative risk 2.3, P less than 0.001) were observed and only 13% of individuals lacked both of these alleles compared with 54% of age- and race-matched controls. A significant association with DRw10 (P = 0.02) was also observed in the DR4/DR1-negative RA group (3/23 patients compared with 0/54 DR4/DR1-negative controls). No novel associations with other DR, DQ, or DP alleles were evident in contrast to some previous studies. The third allelic hypervariable region of the DR beta chain of DRw10 contains a similar amino acid sequence to that found in many DR4 and DR1 molecules. These results extend this correlation and suggest that this susceptibility determinant may account for the HLA-linked susceptibility in 89% of our cases of RA.