ADJUVANT THERAPIES IMPROVING ANTI-REJECTION EFFECTS OF COSTIMULATION BLOCKADE

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 我们获得了20只无特异性病原体的恒河猴肾移植模型。这些动物已经在耶基斯灵长类动物研究中心完成了为期三个月的强制隔离期。在准备肾移植时，我们确定了动物的ABO血型，进行了特异性MHC等位基因分型，进行了移植前混合淋巴细胞反应以验证供体特异性同种异体反应性，并确定了供体和受体对。最初的供体肾切除术定于1月初进行，最初的肾移植将于2月初开始。 已经在体外评价了阿法西普与或不与阿巴西普或贝拉西普的作用。我们发现，灵长类动物效应记忆T细胞的特点是高的CD 2和低的CD 28表达，增加与每一个细胞分裂后同种异体刺激。我们进行了细胞内细胞因子染色，结果表明，CD 8 + CD 2 hiCD 28-细胞显示出最高比例的三重细胞因子产生者（IFN、TNF、IL-2）和细胞毒性效应分子的双重表达者（CD 107 a和颗粒酶B）。在单向混合淋巴细胞反应中，增殖细胞表型为CD 8 + CD 2 hiCD 28-。在体外添加贝拉西普或阿巴西普抑制增殖（1.9- 5倍），添加阿法西普抑制阿坝和贝拉西普抗性增殖。两者合计，这些结果表明，最同种异体反应性T细胞，如增殖和效应分子表达所测量的，绝大多数是CD 28-，并代表一个群体的武装效应，能够逃避共刺激阻断。该共刺激阻断抗性群体的同时高CD 2表达增加了其对阿法赛特的敏感性。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We have obtained 20 specific pathogen free rhesus macaques for renal transplantation. These animals have completed the mandatory three-month quarantine period at the Yerkes Primate Research Center. In preparation for renal transplantation, we have determined the ABO blood types on the animals, performed specific MHC allele typing, performed pre-transplant mixed lymphocyte reactions to verify donor-specific alloreactivity, and determined donor and recipient pairs. Initial donor nephrectomies are scheduled in early January and initial renal transplants will begin in early February. The effects of alefacept with or without abatacept or belatacept have been evaluated in vitro. We found that primate effector memory T cells are characterized by high CD2 and low CD28 expression that increases with each cell division following allo-stimulation. We have performed intracellular cytokine staining and shown that CD8+CD2hiCD28- cells exhibit the highest proportion of both triple cytokine producers (IFN¿, TNF¿, IL-2) and dual expressers of cytotoxic effector molecules (CD107a and granzyme B). In one-way mixed lymphocyte reactions, proliferating cells are phenotypically CD8+CD2hiCD28-. Addition of belatacept or abatacept in vitro inhibited proliferation (1.9- to 5-fold), and addition of alefacept inhibited aba- and belatacept-resistant proliferation. Taken together, these results suggest that the most alloreactive T cells, as measured by both proliferation and effector molecule expression, are overwhelmingly CD28-and represent a population of armed effectors that are able to evade costimulation blockade. The coincident high CD2 expression of this costimulation blockade-resistant population heightens its susceptibility to alefacept.