Van den Bosch Glenn A, Van Gulck Ellen, Ponsaerts Peter, Nijs Griet, Lenjou Marc, Apers Ludwig, Kint Ilse, Heyndrickx Leo, Vanham Guido, Van Bockstaele Dirk R, Berneman Zwi N, Van Tendeloo Viggo F I
Laboratory of Experimental Hematology, Antwerp University (UA), Antwerp University Hospital (UZA), Wilrijkstraat 10, 2650 Edegem, Belgium.
J Immunother. 2006 Sep-Oct;29(5):512-23. doi: 10.1097/01.cji.0000210385.48327.1e.
Recently, it has become obvious that not only CD8 T-cells, but also CD4 T-helper cells are required for the induction of an effective, long-lasting cellular immune response. In view of the clinical importance of cytomegalovirus (CMV) and human immunodeficiency virus (HIV) infection, we developed 2 strategies to simultaneously reactivate viral antigen-specific memory CD4 and CD8 T-cells of CMV-seropositive and HIV-seropositive subjects using mRNA-electroporated autologous CD40-activated B cells. In the setting of HIV, we provide evidence that CD40-activated B cells can be cultured from HAART-naive HIV-1 seropositive patients. These cells not only express and secrete the HIV p24 antigen after electroporation with codon-optimized HIV-1 gag mRNA, but can also be used to in vitro reactivate Gag antigen-specific interferon-gamma-producing CD4 and CD8 autologous T-cells. For the CMV-specific approach, we applied mRNA coding for the pp65 protein coupled to the lysosomal-associated membrane protein-1 to transfect CD40-activated B cells to induce CMV antigen-specific CD4 and CD8 T-cells. More detailed analysis of the activated interferon-gamma-producing CMV pp65 tetramer positive CD8 T-cells revealed an effector memory phenotype with the capacity to produce interleukin-2. Our findings clearly show that the concomitant activation of both CD4 and CD8 (memory) T-cells using mRNA-electroporated CD40-B cells is feasible in CMV and HIV-1-seropositive persons, which indicates the potential value of this approach for application in cellular immunotherapy of infectious diseases.
最近，有一点变得很明显，即诱导有效的、持久的细胞免疫反应不仅需要CD8 T细胞，还需要CD4辅助性T细胞。鉴于巨细胞病毒（CMV）和人类免疫缺陷病毒（HIV）感染的临床重要性，我们开发了两种策略，利用经mRNA电穿孔的自体CD40激活的B细胞，同时重新激活CMV血清阳性和HIV血清阳性受试者的病毒抗原特异性记忆CD4和CD8 T细胞。在HIV感染的情况下，我们提供证据表明，可从未接受高效抗逆转录病毒治疗（HAART）的HIV-1血清阳性患者中培养出CD40激活的B细胞。这些细胞在用密码子优化的HIV-1 gag mRNA进行电穿孔后，不仅表达和分泌HIV p24抗原，还可用于在体外重新激活Gag抗原特异性产生干扰素-γ的自体CD4和CD8 T细胞。对于CMV特异性方法，我们应用编码与溶酶体相关膜蛋白-1偶联的pp65蛋白的mRNA转染CD40激活的B细胞，以诱导CMV抗原特异性CD4和CD8 T细胞。对激活的产生干扰素-γ的CMV pp65四聚体阳性CD8 T细胞进行更详细的分析发现，其具有效应记忆表型，并具有产生白细胞介素-2的能力。我们的研究结果清楚地表明，在CMV和HIV-1血清阳性个体中，使用经mRNA电穿孔的CD40-B细胞同时激活CD4和CD8（记忆）T细胞是可行的，这表明该方法在传染病细胞免疫治疗中的潜在应用价值。
