Analysis of immunoglobulin, complements and CRP levels in serum of captive northern pig-tailed macaques （Macaca leonina）

The northem pig-tailed macaque （NPM, Macaca leonina） has become a widely used animal model in biomedical research. In this study, we measured serum immunoglobulin IgG, IgM, IgA, complement C3, C4 and CRP levels in 3-11 year old captive northem pig-tailed macaques using HITACHI 7600-20 automated chemistry analyzer in order to determine the influences of age and gender on these items. The results showed that serum IgA, IgM, C3 and C4 levels were not correlated with age （P〉0.05）, while serum IgG levels increased progressively with age （r=0.202; P=0.045）. Serum IgG, IgA, IgM and C3 levels were higher in females than in males （P〈0.05）. Moreover, serum C3 concentration was both positively and strongly correlated with that of C4 （r=0.700; P〈0.0001）. This study provides basic serum immunoglobulin and complement data of captive northem pig-tailed macaques, which may prove useful for future breeding efforts and biomedical research.

Flow cytometric characterizations of leukocyte subpopulations in the peripheral blood of northern pig-tailed macaques （Macaca leonina）

Pig-tailed macaques(Macaca nemistrina group) have been extensively used as non-human primate animal models for various human diseases in recent years, notably for AIDS research due to their sensitivity to HIV-1. Northern pig-tailed macaques(M. leonina) are distributed in China and other surrounding Southeast Asia countries. Although northern pig-tailed macaques have been bred on a large scale as experimental animals since 2012, the reference value of normal levels of leukocytes is not available. To obtain such information, 62 blood samples from male and female healthy northern pig-tailed macaques at different ages were collected. The normal range of major leukocyte subpopulations, such as T lymphocytes, B lymphocytes, natural killer(NK) cells, monocytes, and the expression levels of activation or differentiation related molecules(CD38, HLA-DR, CCR5, CD21, IgD, CD80 and CD86) on lymphocytes were analyzed by flow cytometry. The counts of B cells decreased with age, but those of CD8+ T cells and NK cells and the frequency of CD38+HLA-DR+CD4+ T cells were positively correlated with age. The counts of leukocyte subpopulations were higher in males than those in females except for CD4+ T cells. Males also showed higher expression levels of Ig D and CD21 within B cells. This study provides basic data about the leukocyte subpopulations of northern pig-tailed macaques and compares this species with commonly used Chinese rhesus macaques(M. mulatta), which is meaningful for the biomedical application of northern pig-tailed macaques.

Molecular cloning and anti-HIV-1 activities of APOBEC3s from northern pig-tailed macaques(Macaca leonina)

Northern pig-tailed macaques （NPMs, Macaca leonina） are susceptible to HIV-1 infection largely due to the loss of HIV-l-restricting factor TRIM5a. However, great impediments still exist in the persistent replication of HIV-1 in vivo, suggesting some viral restriction factors are reserved in this host The APOBEC3 proteins have demonstrated a capacity to restrict HIV-1 replication, but their inhibitory effects in NPMs remain elusive. In this study, we cloned the NPM A3A-A3H genes, and determined by BLAST searching that their coding sequences （CDSs） showed 99% identity to the corresponding counterparts from rhesus and southern pig-tailed macaques. We further analyzed the anti-HIV-1 activities of the A3A-A3H genes, and found that A3G and A3F had the greatest anti-HIV-1 activity compared with that of other members. The results of this study indicate that A3G and A3F might play critical roles in limiting HIV-1 replication in NPMs in vivo. Furthermore, this research provides valuable information for the optimization of monkey models of HIV-1 infection.

Superior intestinal integrity and limited microbial translocation are associated with lower immune activation in SIVmac239-infected northern pig-tailed macaques(Macaca leonina)

Microbial translocation is a cause of systemic immune activation in HIV/SIV infection. In the present study, we found a lower CD8+ T cell activation level in Macaca leonina (northern pig-tailed macaques, NPMs) than in Macaca mulatta (Chinese rhesus macaques, ChRMs) during SIVmac239 infection. Furthermore, the levels of plasma LPS-binding protein and soluble CD14 in NPMs were lower than those in ChRMs. Compared with ChRMs, SIV-infected NPMs had lower Chiu scores, representing relatively normal intestinal mucosa. In addition, no obvious damage to the ileum or colon epithelial barrier was observed in either infected or uninfected NPMs, which differed to that found in ChRMs. Furthermore, no significant microbial translocation (Escherichia coli) was detected in the colon or ileum of infected or uninfected NPMs, which again differed to that observed in ChRMs. In conclusion, NPMs retained superior intestinal integrity and limited microbial translocation during SIV infection, which may contribute to their lower immune activation compared with ChRMs.

HIV-1 can infect northern pig-tailed macaques(Macaca leonina) and form viral reservoirs in vivo

Viral reservoirs of HIV-1 are a major obstacle for curing AIDS. The novel animal models that can be directly infected with HIV-I will contribute to develop effective strategies for eradicating infections. Here, we inoculated 4 northern pig-tailed macaques （NPM） with the HIV-1 strain HIV-1NL4.3 and moni- tored the infection for approximately 3 years （150 weeks）. The HIV-l-infected NPMs showed transient viremia for about 10 weeks after infection. However, cell-associated proviral DNA and viral RNA persisted in the peripheral blood and lymphoid organs for about 3 years. Moreover, replication-competent HIV-1 could be successfully recovered from peripheral blood mononuclear cells （PBMCs） during long-term infection. The numbers of resting CD4~ T cells in HIV-1 infected NPMs harboring proviruses fell within a range of 2- to 3-1oglo per million cells, and these proviruses could be reactivated both ex vivo and in vivo in response to co-stimulation with the latency-reversing agents JQ1 and prostratin. Our results suggested that NPMs can be infected with HIV-1 and a long-term viral reservoir was formed in NPMs, which might serve as a potential model for HIV-1 reservoir research.

Replication potentials of HIV-1/HSIV in PBMCs from northern pigtailed macaque （Macaca leonina）

The northern pig-tailed macaque （Macaca leonina） has been identified as an independent species of Old World monkey, and we previously found that PBMCs from M. leonina were susceptible to human immunodeficiency virus type 1 （HIV-1）, which may be due to the absence of a TRIM5 protein restricting HIV-1 replication. Here we investigated the infection potentials of six laboratory adapted HIV-1 strains and three primary HIV-1 isolates in PBMCs from M. leonina. The results indicate that these strains are characterized by various but low replication levels, and among which, HIV-INL4-3 shows the highest replication ability. Based on the abundant evidence of species-specific interactions between restriction factors APOBEC3 and HIV/SIV-derived Vif protein, we subsequently examined the replication potentials of v/f-substituted HIV-1 （HSIV） in M. leonina PBMCs. Notably, HSIV-vifmac and stHIV-lsv chimeras, two HIV-1Ni.4-3-derived viruses encoding the viral infectivity factor （Vif） protein from SIVmac239, replicated robustly in cells from M. leonina, which suggests that HSIV could effectively antagonize the antiviral activity of APOBEC3 proteins expressed in cells of M. leonina. Therefore, our data demonstrate that M. leonina has the potential to be developed into a promising animal model for human AIDS.

北平顶猴(Macaca leonina)血液学和血液生化指标参考值的测定与分析

平顶猴是重要的非人灵长类实验动物,广泛应用于艾滋病等重大疾病的动物模型研究。平顶猴分为明打威猴、巽他平顶猴和北平顶猴三个种。我国及周边东南亚国家分布的平顶猴为北平顶猴。该文测定和分析了青幼年(2～4a)和成年(5～10a)北平顶猴的血液学和血液生化指标参考值。结果表明个体性别、年龄和体重等对该指标均有影响。雌性个体的红细胞数、血红蛋白和碱性磷酸酶均低于雄性个体。青幼年个体的白细胞数、淋巴细胞数、单核细胞数、血小板分布宽度、碱性磷酸酶、天冬氨酸转氨酶和胆固醇均高于成年猴,而肌酐和甘油三酯则低于成年猴。个体体重与青年猴的平均血红蛋白含量和肌酐正相关,与成年猴的平均血红蛋白含量、粒细胞百分比、血红蛋白和肌酐正相关,而与成年猴的淋巴细胞数、淋巴细胞百分比、红细胞分布宽度变异系数、天冬氨酸转氨酶和胆固醇负相关。该研究提供的北平顶猴血液学和血液生化指标参考值范围,对其基础研究、疾病诊断和模型建立等具有重要意义。

北平顶猴的规模化饲养与繁殖

北平顶猴是一种珍贵而具有特殊生理特性的灵长类实验动物。介绍了在人工饲养环境下北平顶猴(Macaca mulatta)的驯养、管理、繁殖和疾病防治。综述了养殖场选址、猴房设计建设、设施划分、科学管理、饲养繁殖、个体档案管理以及腹泻、外伤、寄生虫等疾病的预防及治疗措施。

葡萄糖代谢异常导致SIVmac239感染北平顶猴急性期体重变化

目的探讨艾滋病急性期炎症因子、脂类代谢及葡萄糖代谢对体重的影响。方法检测SIVmac239感染北平顶猴急性期炎症因子(TNF-α、IL-6)、脂类代谢(胆固醇、三酰甘油、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇)及葡萄糖代谢参数(胰岛素、胰岛素抵抗指数),比较体重增加组和体重减少组SIV感染北平顶猴之间的差异,运用Pearson相关分析探讨胰岛素抵抗与血浆病毒载量的相关性。结果在SIVmac239感染北平顶猴急性期,炎症因子和血脂四项在体重增加组与体重减少组之间无明显差异;体重增加组胰岛素水平明显下降,体重减少组胰岛素水平明显升高,并出现胰岛素抵抗表现;Pearson分析显示,感染后第5周及第11周胰岛素抵抗指数变化与血浆病毒载量呈正相关。结论 SIVmac239感染急性期,葡萄糖代谢异常可能是导致北平顶猴体重变化的主要原因。

听觉丰容对笼养北豚尾猴行为表达的影响

为了探究笼养动物对听觉丰容的行为响应,以天津动物园笼养北豚尾猴(Macaca leonina)(5只成年,4只未成年)为研究对象,采用扫描取样法,观察研究不同听觉环境(自然音乐、古典音乐和摇滚音乐)对北豚尾猴行为(刻板、理毛、休息和移动)表达的影响.结果表明:①与空白环境相比,自然音乐环境下,群组水平上北豚尾猴的刻板和休息行为增加,理毛和移动行为减少;②古典音乐环境下,北豚尾猴的休息行为增加,刻板和移动行为减少,理毛行为略有减少;③摇滚音乐环境下,北豚尾猴的刻板、理毛、休息和移动行为均减少;④3种音乐环境下,成年北豚尾猴的刻板和理毛行为均多于未成年北豚尾猴,休息和移动行为均少于后者,2个年龄组4种行为的排序均为休息行为>刻板行为>理毛行为>移动行为.综合比较不同听觉环境对北豚尾猴各类行为表达影响的整体效应,认为相比于自然音乐和摇滚音乐,古典音乐更适合作为笼养北豚尾猴听觉丰容的适宜音乐类型.

应用Illumina MiSeq测序技术对腹泻北平顶猴肠道菌群的分析

为探寻可能导致北平顶猴(Macaca leonina)腹泻的菌群及其变化规律,分别采集6只健康和6只腹泻北平顶猴的新鲜粪便,应用Illumina MiSeq高通量测序技术测定其肠道菌群的结构以及腹泻过程中肠道菌群的动态变化。结果表明:健康组肠道菌群的Shannon指数(P=0.015)、Chao 1指数(P=0.014)、ACE指数(P=0.032)较腹泻组高。在门水平上,腹泻组肠道中拟杆菌门(Bacteroidetes)、变形菌门(Proteobacteria)和未鉴定细菌(unidentified Bacteria)的平均相对丰度明显高于健康组,而厚壁菌门(Firmicutes)平均相对丰度低于健康组。在属水平上,腹泻组肠道中的乳杆菌属(Lactobacillus)、普氏菌属(Prevotella)和未鉴定瘤胃球菌属(unidentified Ruminococcaceae)的平均相对丰度明显低于健康组,而链球菌属(Streptococcus)、粪杆菌属(Faecalibacterium)、弯曲杆菌属(Campylobacter)的平均相对丰度明显高于健康组。腹泻与健康北平顶猴肠道菌群丰度具有显著的差异,揭示了这些差异菌群可能是北平顶猴腹泻的重要因素之一。