HLA DRB1*DQB1* haplotype in HTLV-I-associated familial infective dermatitis may predict development of HTLV-I-associated myelopathy/tropical spastic paraparesis

A possible causal association between infective dermatitis and HTLV-I infection was reported in 1990 and confirmed in 1992. We now report familial infective dermatitis (ID) occurring in a 26-year-old mother and her 9-year-old son. The mother was first diagnosed with ID in 1969 at the age of 2 years in the Dermatology Unit at the University Hospital of the West Indies (U.H.W.I.) in Jamaica. The elder of her 2 sons was diagnosed with ID at the age of 3 years, also at U.H.W.I. Both mother and son are HTLV-I-seropositive. A second, younger son, currently age 2 years, is also HTLV-I-seropositive, but without clinical evidence of ID. Major histocompatibility complex (MHC), class II, human leucocyte antigen (HLA) genotyping documented a shared class II haplotype, DRB1*DQB1* (1101-0301), in the mother and her 2 sons. This same haplotype has been described among Japanese patients with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP), and has been associated with a possible pathologically heightened immune response to HTLV-I infection. The presence of this haplotype in these familial ID cases with clinical signs of HAM/TSP may have contributed to their risk for development of HAM/TSP. The unaffected, HTLV-I-seropositive, younger son requires close clinical follow-up. © 1995 Wiley-Liss, Inc.     

Differential diagnosis of HTLV-I-associated myelopathy and multiple sclerosis in Iranian patients

Summary Two Iranian patients with chronic progressive spastic paraparesis and urinary dysfunction were referred to our hospital with the presumptive diagnosis of multiple sclerosis (MS). Routine CSF analysis and magnetic resonance imaging of the two patients were only partially characteristic of MS. Testing for antibodies to human T-cell leukemia virus type I [HTLV-1] in serum using a radioimmune precipitation assay revealed antibodies to HTLV-I in both patients. The infection with HTLV-I was confirmed by polymerase chain reaction (PCR) and liquid hybridization analysis using primers to the tax/rex region and a corresponding probe, demonstrating proviral DNA in peripheral blood mononuclear cells of both patients. On the basis of these findings demonstrating the presence of proviral HTLV-1 DNA in the two Iranian patients, the initial diagnosis of MS was corrected to that of HTLV-I-associated myelopathy (HAM). In contrast, several patients with definite MS (nine from Germany, two from Iran) with a relapsing and remitting form of the disease were tested for HTLV-1 infection by enzyme-linked immunosorbent assay and PCR, which yielded negative results. However, the mother of one HAM patient was found to be infected with HTLV-I. To support an association between HTLV-I infection and CNS disease in the two HAM patients, we analyzed the production of specific IgG antibodies within the CNS based on a simple enzyme immunoassay for viral IgG antibodies in CSF and serum. In the two HAM patients there was significant intrathecal antibody production directed against HTLV-I, but this was not found in any of the samples from MS patients. These findings demonstrate an immune reaction to HTLV-I in the CNS of HAM patients, thus confirming the association of infection and CNS disease. The demonstration of intrathecal HTLV-I antibody production also proved useful for the differential diagnosis of MS or HAM, especially in patients from areas endemic for HTLV-I.Abbreviations DTPA diethylenetriaminepentaacetic acid - ELISA enzyme-linked immunosorbent assay - HAM HTLV-I-associated myelopathy - HTLV-I human T-cell leukemia virus type I - MRI magnetic resonance imaging - MS multiple sclerosis - PBMC peripheral blood mononuclear cells - PCR polymerase chain reaction - RIPA radioimmune precipitation assay - SDS sodium dodecyl sulfate - TSP tropical spastic paraparesis     

A case of HTLV-I associated myelopathy(HAM) in Korea

We report the first Korean case of HTLV-I associated myelopathy (HAM), which was confirmed by Western blot assay of serum and cerebrospinal fluid. Interestingly, the proband's wife was a Japanese Korean who had lived in the endemic area of HAM, Kyushu. Japan, Investigations revealed significantly elevated anti HTLV-I antibody titers in the serum, but not in the CSF, suggesting that she was a carrier of HTLV-I. Considering that the patient had not had a previous blood transfusion, the most common route of HTLV-I, it is likely that the patient was infected by his wife through sexual intercourse. Although previous reports suggest that the transmission of HTLV-I is essentially from man to man or man to woman, our case suggests that woman to man transmission also occurs.     

Tumor necrosis factor, tumor necrosis factor receptors type 1 and 2, lymphotoxin-α, and HLA-DRB1 gene polymorphisms in human T-Cell lymphotropic virus type I associated myelopathy

We studied tumor necrosis factor (TNF), lymphotoxin-α (LT-α), and TNF receptors type 1 (TNFR-1) and type 2 (TNFR-2) gene polymorphisms as well as HLA class II DRB1 alleles in Japanese patients with human T-cell lymphotropic virus type I (HTLV-I) associated myelopathy (HAM) (n = 51), patients with adult T-cell leukemia/lymphoma (ATL) (n = 48), asymptomatic HTLV-I carriers (n = 50), and HTLV-I seronegative, normal controls (n = 112). There were significant differences between HAM patients and normal controls in the distributions of TNF promoter region polymophism at position −857, the LT-α gene NcoI polymorphism, and the T-G substitution in exon 6 of the TNFR-2 gene. The distribution of the NcoI polymorphism of the LT-α gene was also significantly different between HAM patients and asymptomatic HTLV-I carriers. In contrast, we failed to detect any difference in the frequency of DRB1, TNF promoter at position −1031, −863, or the TNFR-1 promoter −383 polymorphism. The results suggest that the TNF/LT-α gene region within the HLA class III of chromosome 6 and the TNFR-2 gene region located on chromosome 1p36 might contribute to susceptibility to HAM, and that aberrant expression or function of these cytokines and the receptor could be involved in the development of HAM.     

An association between serum testosterone level and HLA phenotype

The concentration of testosterone was determined in sera of 122 HLA-typed women. Subsequently the women were classified in the category below or above the mean serum testosterone concentration. When the frequencies of HLA antigens were compared in these two categories of women, it was found that HLA-B5 and HLA-B12 antigens were significantly increased in the category of women with serum testosterone level above the mean concentration (P < 0.01 and P < 0.05, respectively). The frequency of HLA-B8 antigen was significantly decreased in the same category of women (P < 0.05). The comparisons of the mean testosterone values of each HLA antigen and the variance analysis have also shown significant differences between the mean of HLA-B8 antigen and the means of other HLA antigens — HLA-A2, A3, A9, B5, B12 and Bw35. These results gave further conclusive evidence that gene(s) inside HLA region influence either the androgen hormone metabolism itself or cellular sensitivity to hormonal action as it has been presented for congenital adrenal hyperplasia.     

Possible association between HLA antigens and the response to interferon in Japanese patients with chronic hepatitis C

Correlation between the major histocompatibility complex class I antigens (HLA-A, -B and -C) and the elimination from serum of hepatitis C virus in patients with chronic hepatitis C has not been understood. We analyzed HLA phenotypes and their relationship to the efficacy of interferon treatment. Of the 172 patients who were treated with 9 million units of interferon-α2a three times a week for 6 months, 54 patients were responders and 118 patients were non-responders. No significant difference was observed between the 172 patients and 199 healthy subjects with regard to the frequencies of HLA-A, -B and -C antigen phenotypes. However, HLA-B55, B62, CW3 and CW4 frequencies were significantly higher in responders than in non-responders to the interferon treatment. CW4 was found to link with B62, but other phenotypes were independent each other. Patients with HLA B55, B62 and CW3 had a significantly lower viral load, and showed a better response to interferon. These results suggest that HLA system does not have an influence on the evolution towards chronicity of the disease due to hepatitis C virus, but HLA B55, B62 or CW4, and CW3 may be a virus quantity-regulating factors which then affect to response to the interferon treatment, indicating that these HLA antigens in conjunction with a viral peptide is a key target antigen for cytotoxic T lymphocytes in patients with chronic hepatitis C.     

Defective human T-cell lymphotropic virus type I (HTLV-I) provirus in seronegative tropical spastic paraparesis/HTLV-I-associated myelopathy (TSP/HAM) patients

Infection with human T-cell lymphotropic virus type I (HTLV-I) have been associated with the development of the tropical spastic paraparesis/HTLV-I-associated myelopathy (TSP/HAM). We studied the presence of HTLV-I provirus in peripheral blood mononuclear cells (PBMC) from 72 Chilean patients with progressive spastic paraparesis by polymerase chain reaction: 32 seropositive and 40 seronegative cases. We amplified different genomic regions of HTLV-I using primers of 5' ltr, tax, env/tax, pX, pol and env genes. These genes were detected from all seropositive patients. The seronegative patients were negative with 5' ltr, pol, env, and pX primers. However, amplified product of tax and env/tax genes was detected from 16 and four seronegative patients, respectively. Three of them were positive with both genetic regions. The results of this study show that the complete HTLV-I provirus is found in 100% of seropositive cases. In seronegative cases, clinically very similar of seropositive cases, was found only tax gene in 42.5% (17/40) of patients. These results suggest the presence of a defective HTLV-I provirus in some seronegative patients with progressive spastic paraparesis, and suggest a pathogenic role of this truncate provirus for a group of TSP/HAM.     

Reflux of HTLV-I infected lymphocytes from the privileged compartment(s) to peripheral blood flow in patients with HTLV-I-associated myelopathy

 To understand the mechanisms involved in the pathogenesis of human T lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP), three in vivo phenomena which have been observed in the peripheral blood of patients and differing from that in asymptomatic HTLV-I carriers must be taken into consideration: (a) the presence of increased HTLV-I viral load, (b) a higher immune responsiveness against HTLV-I antigens, and (c) biased nucleotide substitutions in the HTLV-I pX region which indicate a decreased selection pressure for viral amino acid changes. We now propose a hypothesis which focuses on the in vivo dynamics of HTLV-I infected lymphocyte migration and which incorporates these features. In addition, the hypothesis assumes the existence of a deviation in immune surveillance for HTLV-I in the central nervous system (CNS) in spite of the presence of frequent specific immune effectors. We suggest that in the active phase of HAM/TSP, accompanied with or following autoaggressive interactions between infected lymphocytes and immunocompetent cells in the CNS, there is a consequential reflux of the infected lymphocytes to the peripheral blood. The reflux of infected cells would be expected to provide peripheral blood with tissue-derived HTLV-I proviruses which have been indulged and propagated in an immune-privileged site. This process would result in and account for the observed increase in viral load and the substitution bias in HTLV-I sequences in the peripheral blood. Received: 31 January 1997 / Accepted: 17 June 1997     

特发性儿童中风患者HLA的相关性研究

目的　探讨与中国北方汉族特发性儿童中风患者相关的HLA抗原及等位基因。方法应用标准微量淋巴细胞毒试验方法及聚合酶链反应 序列特异性探针 (PCR SSO)技术对 43例该病患者进行HLAⅠ类抗原及Ⅱ类等位基因的分型 ,并和 10 7例相匹配的无关健康人进行了对照比较。结果　患者组中HLA B5 1频率为 0 .395 ,而对照组仅为 0 .0 94,相对危险率 (RR)及 χ2 分别为 6 .34和18.94(Pc<0 .0 1)。HLA DRB1 0 80 2频率显著升高 (患者组 0 .2 0 9,对照组 0 .0 2 2 ,RR =11.78,χ2 =13 .5 0 ,Pc=0 .0 0 38)。此外还见到DQA1 0 40 1和DQB1 0 40 2频率也升高 ,校正P值后两组差异仍有显著性 ,二者均与DRB1 0 80 2连锁在同一单倍型中。结论　中国北方汉族特发性儿童中风的发生与HLA B5 1,DRB1 0 80 2显著相关。     

Investigation of the possible association between the HLA antigens and idiopathic thrombocytopenic purpura (ITP)

The etiology of idiopathic thrombocytopenic purpura (ITP), characterized by destruction of platelets, is still poorly understood. Although genetic as well as immunological factors are thought to play a role in the disease pathogenesis, genetic association studies in terms of major histocompatibility complex (MHC) polymorphisms are scarce and discrepant. Results from previous studies suggest that different populations show varying associations with MHC alleles. Since i) there are inconsistencies in HLA associations, and ii) such an association study does not exist for the Indian subcontinent, we carried out sequence specific priming (SSP)-based genotyping of HLA DRB1 alleles in the North Indian population. Data for such studies is available for two East Asian countries, Japan and China, and the association in both cases is different. Further, among the Japanese population too, there are discrepant results. It was therefore important to analyze such an association in the Indian population, belonging to Southern Asia. Our data shows that none of the alleles have any significant association with ITP. Moreover, in contrast to other studies, comparison made between patients who were responsive to steroid therapy against those who were refractory to steroids, also did not show any association of the HLA DRB1 alleles with steroid responsiveness.     

HLA-A位点PCR-SSP基因分型和血清学分型的比较

目的：建立优化ＨＬＡ－Ａ位点ＰＣＲ－ＳＰ分型方法。方法：采用普通扩增仪和Ｅｐｐｅｎｄｏｒｆ管对细胞系ＤＮＡ和３７个健康正常人进行基因分型,血清学检测采用标准淋巴细胞毒试验方法。结果：发现引物浓度和浓度比、ＤＮＡ的纯度及酶的选用,是影响ＨＬＡ－Ａ位点ＰＣＲ－ＳＳＰ分型准确性的重要因素。该方法对３７个标本的基因分型结果与血清学结果相符合。结论：ＰＣＲ－ＳＳＰ方法具有简便准确的优点,可以作为ＨＬＡ－Ａ位     

混合性结缔组织病与HLA抗原的相关性

37例混合性结缔组织病(MCTD)患者的 HLA—A、—B、—DR 抗原分型研究结果表明:在符合 Sharp 标准的 MCTD 患者中,HLA、DR_4、DR_5 抗原频率明显增高,分别为60.9%(RR=4.67,Pc<0.05)和56.5%(RR=4.6,Pc<0.05).而在不符合 Sharp 标准的 MCTD 患者中未发现与 HLA 抗原相关.提示符合 Sharp 标准的 MCTD 患者与不符合该标准的 MCTD 患者的遗传背景存在差异。在研究 MCTD 的不同临床表现型与 HLA 相关性中发现有肌炎表现的 MCTD 患者中 HLA—、DR_4、DR_5抗原频率增高(P<0.05).此外在研究抗 U_1RNP70KD 多肽抗体与 HLA 相关性中发现:抗70KD 阳性同时符合 sharp 标准的患者中 HLA—DR_4、DR_5频率升高(61.9%、56.5%,Pc<0.05),而抗70KD 多肽抗体阳性,但不符合 sharp 标准患者中 HLA—DR_4、DR_5抗原频率并不增高(Pc>0.05).     

Presence of tropical spastic paraparesis/human T-cell lymphotropic virus type 1-associated myelopathy (TSP/HAM)-like among HIV-1-infected patients

Human immunodeficiency virus type 1 (HIV‐1) and human T‐cell lymphotropic virus types 1 and 2 (HTLV‐1 and ‐2) are retroviruses that share similar routes of transmission and some individuals may have a dual infection. These co‐infected subjects may be at increased risk for tropical spastic paraparesis/HTLV‐1‐associated myelopathy (TSP/HAM)‐like. To study the prevalence of tropical spastic paraparesis/HTLV‐1‐associated myelopathy (TSP/HAM) among co‐infected HIV‐1/HTLV‐1 subjects. Since July 1997, our group has been following a cohort to study the interaction of HTLV with HIV and/or hepatitis C virus (HCV), as well as HTLV‐1‐only infected asymptomatic carriers or those already presenting with TSP/HAM. During these 9 years, 296 HTLV‐1‐infected individuals were identified from a total of 538 patients who were referred to our clinic at the Institute of Infectious Diseases “Emílio Ribas,” in São Paulo, Brazil. All subjects were evaluated by two neurologists, blinded to the HTLV status. TSP/HAM diagnosis was based on Kagoshima diagnostic criteria. Results: A total of 38 HIV‐1/HTLV‐1 co‐infected subjects were identified in this cohort: Twenty‐six had already been diagnosed with AIDS and 12 remained asymptomatic. Six of 38 co‐infected subjects (18%) were diagnosed as having TSP/HAM and also AIDS, and for 5 of them TSP/HAM was their first illness. One additional incident case was diagnosed after 2 years of follow‐up. No modifications on HIV‐1 viral load was seen. In contrast, the co‐infected with TSP/HAM‐like group showed higher HTLV‐1 proviral load (505 ± 380 vs. 97 ± 149 copies/10⁴ PBMC, P = 0.012) than asymptomatic co‐infected subjects, respectively. The incidence of myelopathy among HIV‐1/HTLV‐1 co‐infected subjects is probably higher than among patients infected only with HTLV‐1, and related to a higher HTLV‐1 proviral load. Thus, HTLV‐1/2 screening should be done for all HIV‐1‐infected patients in areas where HTLV‐1 infection is endemic. J. Med. Virol. 80:392–398, 2008. © 2008 Wiley‐Liss, Inc.     

HLA and Autoimmunity in Scleroderma (Systemic Sclerosis)

Scleroderma or systemic sclerosis (SSc) has been associated with certain class II antigens of the major histocompatibility complex (MHC), including HLA-DR1, DR2, DR3, DR5, and DR52. In general, these earlier HLA correlations were weak and varied considerably among reporting centers and different ethnic populations. More recently, a variety of disease-specific autoantibodies have been discovered including anticentromere, anti-topoisomerase I, and a variety of anti-nucleolar antibodies. These specificities show little overlap among one another, and each are markers for certain clinical features of SSc.

At the same time, molecular studies of the MHC have provided more accurate methods for defining specific HLA alleles. Now it is becoming clear that certain HLA class II alleles, especially HLA-DQ, are more strongly associated with autoantibody subsets of SSc than with the disease itself. For example, anticentromere antibodies are strongly associated with HLA-DQB1*0501 (DQ5), DQB1*0301 (DQ7) and other DQB1 alleles possessing a glycine or tyrosine residue in position 26 of the outermost domain. Anti-topoisomerase I antibodies occur in SSc patients with HLA-DQB1*0301 (DQ7), DQB1*0302 (DQ8), DQB1*0601 (DQ6 in Japanese), and other DQB1 alleles possessing a tyrosine residue in position 30. HLA-DQ alleles associated with these autoantibodies tend to be in linkage disequilibrium with the HLA-DR specificities previously associated weakly with SSc itself. Rare multiplex families with SSc also show these same HLA haplotypes co-segregating with autoantibody profiles in affected members. Thus, it appears that MHC alleles play a role in affecting the serological expression of SSc, and the implications of these recent findings are discussed.     

Next-Generation Sequencing of the HLA locus: Methods and impacts on HLA typing,population genetics and disease association studies

The human Major Histocompatibility Complex, known as the “Human Leukocyte Antigen (HLA)”, could be defined as a “super locus” (historically called “supergene”) governing the adaptive immune system in vertebrates. It also harbors genes involved in innate immunity. HLA is the most gene-dense, polymorphic and disease-associated region of the human genome. It is of critical medical relevance given its involvement in the fate of the transplanted organs/tissues and its association with more than 100 diseases. However, despite these important roles, comprehensive sequence analysis of the 4 megabase HLA locus has been limited due to technological challenges. Thanks to recent improvements in Next-Generation Sequencing (NGS) technologies however, one is now able to handle the peculiarities of the MHC notably the tight linkage disequilibrium between genes as well as their high degree of polymorphism (and hence heterozygosity). Increased read lengths, throughput, accuracy, as well as development of new bioinformatics tools now enable to efficiently generate complete and accurate full-length HLA haplotypes without phase ambiguities. The present report reviews current NGS approaches to capture, sequence and analyze HLA genes and loci. The impact of these new methodologies on various applications including HLA typing, population genetics and disease association studies are discussed.     

DNA typing of HLA class I (HLA-A) and class II genes (HLA-DR, -DQ and -DP) in Japanese patients with gastric cancer

Gastric cancer is multifactorial disease and several reports have described genetic factors involved in pathogenesis of gastric cancer. Recently, it was reported that HLA class II gene DQB1*0301 was strongly associated with gastric cancer in Caucasian population (20). We performed DNA typing of HLA class I (HLA-A) and HLA class II genes (HLA-DR, DQ and DP) to elucidate the HLA alleles or HLA haplotypes associated with gastric cancer in Japanese population using polymerase chain reaction-sequence-specific oligonucleotide probe analysis in 88 unrelated patients with gastric cancer and 525 unrelated healthy controls. We observed slight difference in frequencies of some HLA alleles and haplotypes between gastric cancer patients and controls; however, after Bonferroni correction, statistical significance was not confirmed. It is possible that environmental factors such as diet cover the contribution of genetic factors to the disease in Japanese population, which has a higher frequency of gastric cancer than do Caucasian populations, most likely due to more exposure to environmental risk factors.     

Extended HLA haplotypes in Japanese homozygous typing cells

Abstract: We have defined extended HLA haplotypes including the HLA class II genes, the non-HLA genes such as TAP1, TAP2 and LMP2, and the (CTG)_n microsatellite repeats within the N0TCH4 gene between DRA and 21OH in 33 Japanese HLA homozygous typing cells (HTC). These conserved haplotypes characterized by unique linkage might be maintained as a result of functional co-operation among them in the antigen presentation pathway. These HTCs can be served as an original and ethnic-specific standard panel, providing useful genetic markers in haplotypic diversity, disease association, and anthropology studies.     

HLA class I alleles in HTLV-1-associated myelopathy and asymptomatic carriers from the Brazilian cohort GIPH

Introduction and objectives  The development of HTLV-1-associated myelopathy (HAM/TSP) in HTLV-1-infected individuals is probably a multi-factor event, in which the immune system plays a crucial role. The efficiency of the host immunity seems to be one of the in vivo determining factors of the proviral load levels and is regulated by genes associated with MHC class I alleles (HLA). Protection or predisposition to HTLV-1-associated diseases according to individual HLA profile was shown in Japanese studies. The present work tested for HLA alleles previously related to protection or susceptibility to HTLV-1-associated myelopathy in a cohort study (GIPH) from Brazil. Methods  A total of 93 HTLV-1-infected individuals participated in the study, as follows: 84 (90.3%) asymptomatic and 9 (9.7%) with HAM/TSP. Alleles related to protection (A*02, Cw*08) and susceptibility (B*07, Cw*08 and B*5401) were tested by the PCR-SSP method. Results  Allele A*02 was more frequent in the asymptomatic group and in its absence, Cw*07 was correlated with HAM/TSP (P = 0.002). Allele B*5401 was not present in the Brazilian population. Alleles B*07 and Cw*08 were not different between the groups Discussion  The presence of HLA-A2 elicits a stronger cytotoxic response, which is involved in the HTLV-1 proviral load reduction. This study confirmed a tendency of this allele to protect against HAM-TSP. Therefore, A*02 might be of interest for researches involved with HTLV-1 vacine.