Analysis of primary immunodeficiency syndrome with low levels of antibody: diagnosis and treatment

原发性免疫缺陷综合征低抗体分析：诊断与治疗

• 批准号: 13470162
• 负责人: KOMIYAMA Atsushi
• 金额: $ 5.57万
• 依托单位: Shinshu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470162/
• 关键词: Plasma cells; Memory B cells; CD27; immunodeficiency; Naive B cells; 抗体欠乏; 重症複合免疫不全症; 分類不能型免疫不全症; 遺伝子治療

To produce antibodies, the differentiation of B cells into antibody-secreting cells, plasma cells, is required. We describe that a ligation of CD27, which belongs to the tumor necrosis factor receptor (TNFR) family and is memory marker of B cells, is memory B-cell marker and yields crucial signals that positively control the entry of B cells into the pathway to plasma cells. On the basis of this finding, we investigated B-cell functions in primary immunodeficiencies with low levels of immunoglobulins in the serum, and performed new approach about the diagnosis and the treatment in primary immunodeficiencies.we describe a novel mutation of genomic exon 3 deletion, resulting in exon 3 and 4 m RNA skipping, of Artemis gene found in 4 Japanese patients of 4 unrelated families with severe combined immunodeficiency with absence of immunoglobulin and increased cellular radiosensitivity (RS-SCID) . The heterozygous genornic exon 3 deletion was also found in their parents studied. This report i … More s the first study of Artemis deficiency in East Asia and suggests that the Artemis gene mutation is peculiar to Japanese.In regarding common variable immunodeficiency, The molecular basis of common variable immunodeficiency (CVID) is unknown. To assess humoral immunity in CVID, we selected 24 patients with early or late onset of disease. X-linked agammaglobulinemia (XLA), X-linked hyper-IgM syndrome (XHIM) and non-XHIM were excluded based on clinical phenotype, assessment of the immune response, presence of Bruton's tyrosine kinase (Btk) in monocytes or platelets and normal expression of CD40 ligand by activated T cells. The number of circulating B cells was within normal range or reduced. IgD^- CD27^+ memory B-cells were markedly reduced or absent in all 24 patients and IgD^+ CD27^+ B cells were diminished in 8 patients. Circulating B cells from all six patients examined, including CVID patients with IgD^+ CD27^+ cells, failed to undergo somatic hypermutation in immunoglobulin variable (V)-region genes, similar to cord blood B cells. B cells from CVID patients produced IgM and IgG, but not IgA upon the engagement of Ig receptor and CD40 in the presence of IL-2 and IL-10. B cells from all but 5 patients secreted IgE when stimulated by CD40 crosslinking in the presence of JL-4. The observation of defective memory B cells with abnormal cell marker expression and function demonstrates that naive CVID B cells including those expressing IgD^+ CD27^+, in analogy to cord blood and hyper IgM syndrome B cells, may be responsible for their failure to differentiate into plasma cells and to produce high affinity antibodies of different isotypes.In the patients with X-linked hyper-IgM syndrome (XHIM), costimulation of mononuclear cells from most of the patients induced no to low levels of class switching from IgM to IgG and IgA with Staphylococcus aureus Cowan strain (SAC) plus IL-2 or anti-CD40 mAb (anti-CD40) plus IL-10. Measurable levels of IgE were secreted in some of the patients after stimulation with anti-CD40 plus IL-4. Costimulation with SAC plus IL-2 plus anti-CD40 plus IL-10 yielded secretion of significant levels of IgG in addition to IgM, but not IgA. The most striking finding was that peripheral blood B cells from all of the six patients were composed of only IgD+ CD27- and IgD+ CD27+ B cells; IgD- CD27+ memory B cells were greatly decreased. IgD+ CD27+ B cells from an XHIM patient produced IgM predominantly. Our data indicate that the low response of IgG production in XHIM patients is due to reduced numbers of IgD- CD27+ memory B cells . However, the IgG production can be induced by stimulation of immunoglobulin receptors and CD40 in cooperation with such cytokines as IL-2 and IL-10 in vitro. Less

为了产生抗体，需要将B细胞分化为分泌抗体的细胞，即浆细胞。我们描述了属于肿瘤坏死因子受体（TNFR）家族并且是B细胞的记忆标志物的CD 27的连接是记忆B细胞标志物并且产生积极控制B细胞进入浆细胞通路的关键信号。在此基础上，我们研究了血清免疫球蛋白水平低的原发性免疫缺陷患者的B细胞功能，并对原发性免疫缺陷的诊断和治疗进行了新的探索。我们描述了一种新的基因组外显子3缺失突变，导致外显子3和4 mRNA跳跃，Artemis基因在4个日本患者的4个无关的家庭，严重的联合免疫缺陷缺乏免疫球蛋白和细胞放射敏感性增加（RS-SCID）。在所研究的父母中也发现了杂合性外显子3缺失。这份报告i ...更多信息 本研究首次在东亚地区发现Artemis缺乏症，提示Artemis基因突变是日本人特有的。为了评估CVID中的体液免疫，我们选择了24例早发或迟发疾病患者。根据临床表型、免疫应答评估、单核细胞或血小板中存在布鲁顿酪氨酸激酶（Btk）以及活化T细胞正常表达CD 40配体，排除X连锁无丙种球蛋白血症（XLA）、X连锁高IgM综合征（XHIM）和非XHIM。循环B细胞数量在正常范围内或减少。所有24例患者的IgD^-CD 27 ^+记忆B细胞均显著减少或缺失，8例患者的IgD^+ CD 27 ^+ B细胞减少。所有6例患者的循环B细胞，包括IgD^+ CD 27 ^+细胞的CVID患者，均未发生免疫球蛋白可变区（V）基因的体细胞高突变，与脐带血B细胞相似。CVID患者的B细胞在IL-2和IL-10存在下与IG受体和CD 40结合时产生IgM和IgG，但不产生伊加。当在JL-4存在下通过CD 40交联刺激时，除5名患者外，所有患者的B细胞均分泌IgE。通过观察具有异常细胞标志物表达和功能的缺陷性记忆B细胞，证实幼稚CVID B细胞，包括表达IgD^+ CD 27 ^+的细胞，类似于脐带血和高IgM综合征B细胞，可能是它们不能分化为浆细胞和产生不同同种型的高亲和力抗体的原因。用金黄色葡萄球菌科万菌株（SAC）加IL-2或抗-CD 40 mAb（抗-CD 40）加IL-10共刺激来自大多数患者的单核细胞诱导无至低水平的从IgM到IgG和伊加的类别转换。在用抗CD 40加IL-4刺激后，一些患者分泌了可测量水平的IgE。用SAC加IL-2加抗CD 40加IL-10共刺激产生除了IgM之外的显著水平的IgG分泌，但不产生伊加。最引人注目的发现是来自所有6名患者的外周血B细胞仅由IgD+ CD 27-和IgD+ CD 27 + B细胞组成; IgD-CD 27+记忆B细胞显著减少。来自XHIM患者的IgD+ CD 27 + B细胞主要产生IgM。我们的数据表明，XHIM患者中IgG产生的低应答是由于IgD-CD 27+记忆B细胞数量减少。然而，IgG的产生可以通过刺激免疫球蛋白受体和CD 40与细胞因子如IL-2和IL-10在体外协同诱导。少

项目成果

Uehara Y, Kikuchi K, Nakamura T, Nakama H, Agematsu K, Kawakami Y, Maruchi N, Totsuka K: "H202 Produced by Viridans Group Streptococci May Contribute to Inhibition of Methicillin-Resistant Staphylococcus aureus Colonization of Oral Cavities in Newborns"Cl

Uehara Y、Kikuchi K、Nakamura T、Nakama H、Agematsu K、Kawakami Y、Maruchi N、Totsuka K：“Viridans 群链球菌产生的 H202 可能有助于抑制新生儿口腔中耐甲氧西林金黄色葡萄球菌的定植”Cl

Gombart A.F., Shiohara M., Kwok S.H., Agematsu K., Komiyama A., Koeffier H.P.: "Neutrophil-specific granule deficiency: homozygous recessive inheritance of a frameshift mutation in the gene encoding transcription factor CCAAT/enhancer binding protein-epsi

Gombart A.F.、Shiohara M.、Kwok S.H.、Agematsu K.、Komiyama A.、Koeffier H.P.：“中性粒细胞特异性颗粒缺陷：编码转录因子 CCAAT/增强子结合蛋白-epsi 的基因中移码突变的纯合隐性遗传

nagumo H, Agematsu K, Kobayashi N, Shinozaki S, Hokibara S, Nagase H, Takamoto M, Yasui K, Sugane S, Komiyama A: "Different process of class switching and somatic hypermutation : a novel analysis by CD27^- naive B cells"Blood. 15. 567-575 (2002)

nagumo H, Agematsu K, Kobayashi N, Shinozaki S, Hokibara S, Nagase H, Takamoto M, Yasui K, Sugane S, Komiyama A：“类别转换和体细胞超突变的不同过程：CD27^-幼稚 B 细胞的新分析

Uehara Y, Kikuchi K, Nakamura T, Nakama H, Agematsu K, Kawakami Y, Maruchi N, Totsuka K: "H2O2 Produced by Viridans Group Streptococci May Contribute to Inhibition of Methicillin-Resistant Staphylococcus aureus Colonization of Oral Cavities in Newborns"Cl

Uehara Y、Kikuchi K、Nakamura T、Nakama H、Agematsu K、Kawakami Y、Maruchi N、Totsuka K：“Viridans 群链球菌产生的 H2O2 可能有助于抑制新生儿口腔中耐甲氧西林金黄色葡萄球菌的定植”Cl

Nagumo H, Agematsu K, Kobayashi N, Shinozaki S, Hokibara S, Nagase H, Takamoto M, Yasui K, Sugane S, Komiyama A: "Different process of class switching and somatic hypermutation ; a novel analysis by CD27^-naive B cells"Blood. 99. 567-575 (2002)

Nagumo H、Agematsu K、Kobayashi N、Shinozaki S、Hokibara S、Nagase H、Takamoto M、Yasui K、Sugane S、Komiyama A：“类别转换和体细胞超突变的不同过程；CD27^-naive B 细胞的新分析