HELPER T-CELL SUBSETS AND MUCOSAL IMMUNITY

辅助 T 细胞亚群和粘膜免疫

• 批准号: 2004077
• 负责人: HIROSHI KIYONO
• 金额: $ 11.53万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-11-01 至 1997-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2004077
• 关键词: Macaca mulatta; cell mediated cytotoxicity; cell type; cellular immunity; cholera toxin; cytotoxic T lymphocyte; disease /disorder model; drug delivery systems; enzyme linked immunosorbent assay; glycoproteins; helper T lymphocyte; immunization; immunologic memory; interferon gamma; interleukin 10; interleukin 2; interleukin 4; interleukin 8; microcapsule; mucosal immunity; polymerase chain reaction; recombinant proteins; simian immunodeficiency virus; vector vaccine; viral vaccines; virus protein

Recent studies in the murine system have shown that CD4+ T helper (Th) cells can often be subdivided based upon profiles of cytokines produced, and convincing evidence is now at hand for Th cell subsets in humans with autoimmune, allergic and infectious diseases. Two subsets, Th1 and Th2 (type 1 and type 2) are best characterized and type 1 Th cells produce interleukin-2 (IL-2), interferon gamma (IFN-gamma) and TNF-beta for cell- mediated immunity (CMI) in order to protect against intracellular parasites. Type 2 Th cells produce IL-4, IL-5, IL-6 and IL-10 and upregulate IgG subclass, IgE and IgA antibody responses. Important recent studies have shown that after HIV infection, the progression to AIDS is characterized by loss of Th1 cells and with increases in Th2 cells which are associated with well documented increases in serum IgG and IgA levels. Our own studies have recently shown that oral immunization induced mainly Th2-type cells for regulation of mucosal secretory IgA (S-IgA) responses. Thus, we have postulated that mucosal vaccines should induce Th2 cells for optimal S-IgA responses and Th1-type cells for CMI and help for CD8+ cytotoxic T lymphocyte (CTL) responses. We have further hypothesized that SIV infection results in loss of TH1 cells in systemic tissues; however mucosal CD4+ Th2 cell responses may remain functional. Further, this hypothesis would suggest that SIV/HIV vaccines should optimize induction of Th2 cells for mucosal antibodies as well as type 1 cells for systemic CMI and CTL responses. To test this, our grant has been divided into two parts. In the first part, we will assess Th1 and Th2 cell imbalances in both systemic and mucosa-associated tissues of SIV infected monkeys. We will use single-cell assays, i.e., enzyme-linked immunospots (ELISPOTs) to assess Th1 (IFN-gamma and IL-2) and Th2(IL04 and IL-10)cytokines as well as RNA protection assay (RPA) and reverse transcriptase (RT)-PCR for analysis of cytokine mRNA. The first specific aim will focus on peripheral blood CD4+T cells for memory (recall) type 1 or type 2 responses to standard vaccines and to mitogen (PHA). The second specific aim will continue to assess Th1/Th2 imbalances in SIV infection, but will emphasize CD4+ Th cell responses to recall vaccine, to PHA and to SIV components with T lymphocytes isolated from both mucosa-associated and systemic lymphoid tissues. The overall emphasis in the second part of this grant will be to assess optimal Th1 and Th2 cell responses following deliberate mucosal immunization with whole SIV or its components (rgp130 and gp27). We plan state-of-the-art mucosal immunization strategies using cholera toxin (CT) and with SIV gp130 components conjugated to CT-B as adjuvant as well water- based microspheres and recombinant bacteria (rSalmonella tryphimurium and rVibrio cholerae) for mucosal delivery of SIV components. In specific aim 3, we will use CT as adjuvant to optimize mucosal immune responses including induction of type 1 and especially type 2 CD4+ responses. In the final aim, we plan to compare other delivery systems, i.e., water-based microspheres and recombinant bacteria for mucosal delivery. This effort is part of a Collaboration with four other groups with expertise in heterosexual (CRPRC) and homosexual (Guy's/CAMR) SIV infection and immunity, with experience in mucosal vector systems (VRI) and with unique models to study S-gA antibody function, in vitro (CWR). Together we can approach the goal of inducing effective mucosal and systemic immunity fy using appropriate cellular and molecular techniques developed for rhesus macaques.

最近在小鼠系统中的研究表明，CD 4 + T辅助细胞（Th） 细胞通常可以基于产生的细胞因子的分布而细分， 现在有令人信服的证据表明， 自身免疫性、过敏性和感染性疾病。 两个亚群，Th 1和Th 2 (type 1型和2型）的特征最好，1型Th细胞产生 白细胞介素-2（IL-2）、干扰素γ（IFN-γ）和TNF-β用于细胞- 介导的免疫（CMI），以防止细胞内 寄生虫 2型Th细胞产生IL-4、IL-5、IL-6和IL-10， 上调IgG亚类、IgE和伊加抗体应答。 最近取得的重大 研究表明，艾滋病毒感染后， 其特征在于Th 1细胞的损失和Th 2细胞的增加， 与血清IgG和伊加水平的增加有关。 我们自己的研究最近表明，口服免疫主要诱导 调节粘膜分泌型伊加（S-IgA）应答的Th 2型细胞。 因此，我们假设粘膜疫苗应该诱导Th 2细胞， 最佳的S-IgA反应和Th 1型细胞用于CMI和帮助CD 8 + 细胞毒性T淋巴细胞（CTL）应答。 我们进一步假设， SIV感染导致全身组织中TH 1细胞的损失;然而， 粘膜CD 4 + Th 2细胞应答可能保持功能性。 此外，这 一种假说认为，SIV/HIV疫苗应优化诱导 粘膜抗体的Th 2细胞以及系统性CMI的1型细胞 CTL反应。 为了验证这一点，我们的补助金被分为两部分 零件. 在第一部分中，我们将评估Th 1和Th 2细胞失衡， SIV感染猴的全身和粘膜相关组织。 我们 将使用单细胞测定，即，酶联免疫斑点（ELISPOTs）， 评估Th 1（IFN-γ和IL-2）和Th 2（IL-04和IL-10）细胞因子以及 RNA保护试验（RPA）和逆转录酶（RT）-PCR用于分析 细胞因子mRNA。 第一个具体目标将集中在外周血 CD 4 +T细胞用于记忆（回忆）1型或2型对标准 疫苗和促分裂原（PHA）。 第二个具体目标将继续 评估SIV感染中Th 1/Th 2失衡，但将强调CD 4 + Th细胞 对召回疫苗、PHA和SIV组分的反应， 从粘膜相关和全身淋巴细胞中分离的淋巴细胞 组织中 该补助金第二部分的总体重点将是 评估在故意粘膜刺激后的最佳Th 1和Th 2细胞应答 用完整SIV或其组分（rgp 130和gp 27）免疫。 我们计划 使用霍乱毒素（CT）的最新粘膜免疫策略 和与CT-B缀合的SIV gp 130组分作为佐剂以及水- 的微球和重组细菌（鼠伤寒沙门氏菌和 rVibrio cholesterol）用于粘膜递送SIV组分。 具体目标 3、以CT为佐剂优化黏膜免疫应答 包括诱导1型和特别是2型CD 4+应答。 在 最后，我们计划比较其他的输送系统，即，水基 用于粘膜递送的微球和重组细菌。 这一努力是 与其他四个具有以下专门知识的小组合作的一部分： 异性恋（CRPRC）和同性恋（Guy's/CAMR）SIV感染， 免疫，具有粘膜载体系统（VRI）的经验， 体外研究S-gA抗体功能的模型（CWR）。 我们可以一起 达到诱导有效的粘膜和全身免疫的目的， 使用为恒河猴开发的适当的细胞和分子技术 猕猴

项目成果

Nasal immunization of nonhuman primates with simian immunodeficiency virus p55gag and cholera toxin adjuvant induces Th1/Th2 help for virus-specific immune responses in reproductive tissues.

用猿猴免疫缺陷病毒 p55gag 和霍乱毒素佐剂对非人灵长类动物进行鼻免疫，可诱导 Th1/Th2 帮助生殖组织中的病毒特异性免疫反应。

• 发表时间: 1998
• 期刊: Journal of immunology (Baltimore, Md. : 1950)
• 作者: Imaoka,K;Miller,CJ;Kubota,M;McChesney,MB;Lohman,B;Yamamoto,M;Fujihashi,K;Someya,K;Honda,M;McGhee,JR;Kiyono,H
• 通讯作者: Kiyono,H

Selected Th1 and Th2 cytokine mRNA expression by CD4(+) T cells isolated from inflamed human gingival tissues.

从发炎的人牙龈组织中分离的 CD4(+) T 细胞表达选定的 Th1 和 Th2 细胞因子 mRNA。

• DOI: 10.1111/j.1365-2249.1996.tb08297.x
• 发表时间: 1996
• 期刊: Clinical and experimental immunology
• 影响因子: 4.6
• 作者: Fujihashi,K;Yamamoto,M;Hiroi,T;Bamberg,TV;McGhee,JR;Kiyono,H
• 通讯作者: Kiyono,H

gamma/delta T cell-deficient mice have impaired mucosal immunoglobulin A responses.

• DOI: 10.1084/jem.183.4.1929
• 发表时间: 1996-04-01
• 期刊: The Journal of experimental medicine
• 作者: Fujihashi K;McGhee JR;Kweon MN;Cooper MD;Tonegawa S;Takahashi I;Hiroi T;Mestecky J;Kiyono H
• 通讯作者: Kiyono H

Oral immunization with simian immunodeficiency virus p55gag and cholera toxin elicits both mucosal IgA and systemic IgG immune responses in nonhuman primates.

口服免疫缺陷病毒 p55gag 和霍乱毒素可在非人灵长类动物中引发粘膜 IgA 和全身 IgG 免疫反应。

• 发表时间: 1997
• 期刊: Journal of immunology (Baltimore, Md. : 1950)
• 作者: Kubota,M;Miller,CJ;Imaoka,K;Kawabata,S;Fujihashi,K;McGhee,JR;Kiyono,H
• 通讯作者: Kiyono,H

Mucosal Th1- versus Th2-type responses for antibody- or cell-mediated immunity to simian immunodeficiency virus in rhesus macaques.

恒河猴中针对猿猴免疫缺陷病毒的抗体或细胞介导免疫的粘膜 Th1 型与 Th2 型反应。

• DOI: 10.1086/314807
• 发表时间: 1999
• 期刊: The Journal of infectious diseases.
• 作者: McGhee,JR;Kiyono,H;Kubota,M;Kawabata,S;Miller,CJ;Lehner,T;Imaoka,K;Fujihashi,K
• 通讯作者: Fujihashi,K