An M2 Rather than a TH2 Response Contributes to Better Protection against Latency Reactivation following Ocular Infection of Naive Mice with a Recombinant Herpes Simplex Virus 1 Expressing Murine Interleukin-4
An M2 Rather than a TH2 Response Contributes to Better Protection against Latency Reactivation following Ocular Infection of Naive Mice with a Recombinant Herpes Simplex Virus 1 Expressing Murine Interleukin-4
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DOI：
10.1128/jvi.00051-18
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发表时间：
2018-05-01
影响因子：
5.4
通讯作者：
Ghiasi, Homayon
中科院分区：
文献类型：
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作者：
Lee, Dhong Hyun;Ghiasi, Homayon
我们先前发现，在眼部感染单纯疱疹病毒1型(HSV-1)的小鼠中，注射集落刺激因子1(CSF-1)改变巨噬细胞对M2反应的极化比注射干扰素(IFN-Gamma)改变M1极化更有效地减少初发和潜伏感染。细胞因子可协同调节巨噬细胞和辅助性T细胞(T-H)应答，其中白细胞介素4(IL-4)诱导2型T-H(T(H)2)，M2应答，干扰素-γ诱导T(H)1应答，M1应答。现在，我们通过比较感染和重组HSV-1的效果来区分这些免疫区段在预防潜伏期重新激活和角膜瘢痕形成中的作用。在重组HSV-1中，潜伏期相关转录本(LAT)基因被IL-4(HSV-IL-4)或干扰素-γ(HSV-γ)基因取代，并以亲本(LAT-阴性)病毒感染为对照。体外对腹膜巨噬细胞的分析表明，用IL-4或干扰素-γ基因取代LAT不会影响病毒的感染性，并适当地促进极化。眼部感染HSV-IL-4的小鼠对角膜疤痕的保护作用显著高于HSV-干扰素-伽马或亲本病毒感染的小鼠。3组小鼠眼部和三叉神经节(TG)的原代病毒复制水平相似，但HSV-IL-4感染组小鼠眼部GC(+)细胞数在感染后第5天明显低于HSV-干扰素-γ组或亲本病毒组。单纯疱疹病毒-IL-4和单纯疱疹病毒干扰素-γ感染的小鼠的潜伏期和外植体激活均低于亲本病毒感染的小鼠，潜伏期最低的水平与单纯疱疹病毒-IL-4感染有关。潜伏期较长与CD8、PD-1和干扰素-伽马mRNA水平较高相关，而潜伏期缩短和T细胞耗竭与TG中较低的GC(+)表达相关。在所有眼部感染的小鼠中，与未耗尽的小鼠相比，巨噬细胞耗竭增加了潜伏期水平，其中HSV-IL-4组的巨噬细胞耗竭使潜伏期增加了3000倍以上。我们的结果表明，在HSV-1感染中，将固有巨噬细胞免疫反应转移到M2，而不是M1，将提高对潜伏期、重新激活和眼部疾病的保护。眼部HSV-1感染是美国最常见的严重病毒性眼部感染之一，也是病毒致盲的主要原因。由于在三叉神经节建立潜伏感染会导致反复感染，并与角膜瘢痕形成相关，因此预防潜伏再激活是一个主要的治疗目标。众所周知，缺乏潜伏期相关转录本(LAT)可以减少潜伏期重新激活。在这里，我们证明了表达IL-4(T(H)2/M2应答的诱导剂)或干扰素-γ(T(H)1/M1应答的诱导剂)的重组HSV-1替代LAT进一步缩短了潜伏期，其中HSV-IL-4显示出最高的整体保护效果。在幼稚的小鼠中，这种更高的保护效果是由先天免疫反应而不是获得性免疫反应介导的。虽然M1和M2巨噬细胞的反应都是保护性的，但通过表达IL-4使巨噬细胞向M2反应转变在缩短眼部HSV-1潜伏期重新激活方面更有效。
We found previously that altering macrophage polarization toward M2 responses by injection of colony-stimulating factor 1 (CSF-1) was more effective in reducing both primary and latent infections in mice ocularly infected with herpes simplex virus 1 (HSV-1) than M1 polarization by gamma interferon (IFN-gamma) injection. Cytokines can coordinately regulate macrophage and T helper (T-H) responses, with interleukin-4 (IL-4) inducing type 2 T-H (T(H)2) as well as M2 responses and IFN-gamma inducing T(H)1 as well as M1 responses. We have now differentiated the contributions of these immune compartments to protection against latency reactivation and corneal scarring by comparing the effects of infection with recombinant HSV-1 in which the latency-associated transcript (LAT) gene was replaced with either the IL-4 (HSV-IL-4) or IFN-gamma (HSV-IFN-gamma) gene using infection with the parental (LAT-negative) virus as a control. Analysis of peritoneal macrophages in vitro established that the replacement of LAT with the IL-4 or IFN-gamma gene did not affect virus infectivity and promoted polarization appropriately. Protection against corneal scarring was significantly higher in mice ocularly infected with HSV-IL-4 than in those infected with HSV-IFN-gamma or parental virus. Levels of primary virus replication in the eyes and trigeminal ganglia (TG) were similar in the three groups of mice, but the numbers of gC(+) cells were lower on day 5 postinfection in the eyes of HSV-IL-4-infected mice than in those infected with HSV-IFN-gamma or parental virus. Latency and explant reactivation were lower in both HSV-IL-4- and HSV-IFN-gamma-infected mice than in those infected with parental virus, with the lowest level of latency being associated with HSV-IL-4 infection. Higher latency correlated with higher levels of CD8, PD-1, and IFN-gamma mRNA, while reduced latency and T-cell exhaustion correlated with lower gC(+) expression in the TG. Depletion of macrophages increased the levels of latency in all ocularly infected mice compared with their undepleted counterparts, with macrophage depletion increasing latency in the HSV-IL-4 group greater than 3,000-fold. Our results suggest that shifting the innate macrophage immune responses toward M2, rather than M1, responses in HSV-1 infection would improve protection against establishment of latency, reactivation, and eye disease.IMPORTANCE Ocular HSV-1 infections are among the most frequent serious viral eye infections in the United States and a major cause of virus-induced blindness. As establishment of a latent infection in the trigeminal ganglia results in recurrent infection and is associated with corneal scarring, prevention of latency reactivation is a major therapeutic goal. It is well established that absence of latency-associated transcripts (LATs) reduces latency reactivation. Here we demonstrate that recombinant HSV-1 expressing IL-4 (an inducer of T(H)2/M2 responses) or IFN-gamma (an inducer of T(H)1/M1 responses) in place of LAT further reduced latency, with HSV-IL-4 showing the highest overall protective efficacy. In naive mice, this higher protective efficacy was mediated by innate rather than adaptive immune responses. Although both M1 and M2 macrophage responses were protective, shifting macrophages toward an M2 response through expression of IL-4 was more effective in curtailing ocular HSV-1 latency reactivation.