Study on HLA-B27 binding antigenic peptides in the patients with ankylosing spondylitis.

1.采用聚合酶链式反应-序列特异性寡核苷酸探针法(PCR-SSOP)检测日本脊柱关节病患者和健康人的HL A-B27亚型。日本人以B^&lt；**&gt；2704亚型为主，其次为B^&lt；**&gt；2705亚型。这两种亚型都与疾病有关。B^&lt；**&gt；2704与A^&lt；**&gt；1102之间存在连锁不平衡。#2.采用单链构象多态(Single-strand Configuration Polysis，PCSSCP)技术，对HLAB27阳性日本患者和健康体检者的MCHA等位基因进行了检测。HLA-B^&lt；**&gt；2704与MICA 010密切相关，而HLA-B^&lt；**&gt；2705与MICA 007处于强连锁不平衡状态。#3.比较了同卵双生子B淋巴母细胞系(B-LCLS)与强直性脊柱炎的关系。从每个细胞系中都检测到九种多肽。测定了其中一种多肽的氨基酸序列。在患者和对照组之间没有观察到质的差异。还试图鉴定29例患者外周血白细胞中洗脱的HLA-B27结合多肽，但没有获得足够数量的多肽用于分析。#4.测定血清中可溶性HLAI类抗原的水平。系统性红斑狼疮(SLE)患者sHLAI类分子显著升高。SLE患者血清sHLAI类分子浓度与疾病活动性呈显著正相关。SHLAI类分子主要来源于患者体内的蛋白水解性切割和选择性剪接。

#1. HLA-B27 subtypes of Japanese patients with spondyloarthropathies and healthy individuals were determined using polymerase chain reaction followed by sequence-specific oligonucleotide probing (PCR-SSOP). The dominant HLA-B27 subtype among Japanese was shown to be HLA-B^<**>2704, followed by B^<**>2705. Both subtypes were associated with the diseases. The linkage disequilibrium between B^<**>2704 and A^<**>1102 was also demonstrated.#2. MCHA (MHC class I chain-related gene A) alleles of HLA-B27 positive Japanese patients with AS as well as healthy individuals were determined using PCR-SSCP (single-strand conformation polymorphism) technique. HLA-B^<**>2704 was tightly associated with MICA 010, while HLA-B^<**>2705 was in strong linkage disequilibrium with MICA 007.#3. The peptides eluted from HLA-B27 molecules of B lymphoblastoid cell lines (B-LCLs) from identical twins discordant for ankylosing spondilitis were compared. Nine peptides were detected from each of the cell lines. Amino acid sequence of one the peptides was determined. No qualitative difference was observed between the patient and the control. Attempts were also made to identify HLA-B27-binding peptides eluted from pooled peripheral blood leukocytes from 29 patients, however, sufficient quantity of peptides for the analysis was not obtained.#4. Serum level of soluble HLA-class I antigens (sHLA-class I) was determined. sHLA-class I was significantly elevated in patients with systemic lupus erythematosus (SLE). Significant correlation of sHLA-class I concentration and disease activity of SLE was observed. sHLA-class I was shown to be derived mainly from proteolytic cleavage and alternative splicing in patients.