In vitro aging of macaque adherent cells: similar pattern of cellular aging between human and macaque

To explore new models for human cellular aging as well as to evaluate aging of the macaques, profiles of cellular aging in macaques were studied. Adherent cells were obtained from five Japanese macaques (Macaca fuscata), 14 long-tailed macaques (Macaca fascicularis), two bonnet monkeys (Macaca radiata) and a rhesus monkey (Macaca mulatta). A total of 35 cultures were performed and cell morphology, doubling time, telomere length and telomerase activity were studied. They were classified into three groups; group I: cell strains with a definite replicative life-span (-41 PDLs) (presence of M1), group II: cell strains with a limited extension of replicative life-span (79-106 PDLs) with p53 mutation(s) (presence of M2), and group III: a cell strain with an indefinite replicative life-span (>150 PDLs) with characteristics of transformation. Except for the last group, telomerase activity was not observed. Macaque cells demonstrated three chronological patterns comprising both human and rodent patterns, however, presence of the two limits of proliferation in vitro grants macaque cells to be more appropriate than rodents in both studying human aging and oncogenesis.     

The modelling of hepatitis A in macaques]

Characteristics of experimental hepatitis A in Macaca fascicularis and M. mulatta produced with HAV, strain MP, isolated from M. mulatta in an outbreak of spontaneous hepatitis are presented. The HAV-MP strain induced the disease in all the animals used in the experiment. The infection was manifest, with all virological, serological, biochemical and morphological features typical of hepatitis A. In M. mulatta, the process was protracted, with virus persistence in feces for at least 4 months. Modeling of hepatitis A in Macaca monkeys using HAV-MP strain may be used for the study of the pathogenesis and trials of immune preparations.     

The immune structure and specific laboratory indices of acute hepatitis A in lower monkeys of the Sukhumi Nursery

The study of the rate of occurrence of hepatitis A (HA) markers among monkeys, both newly arriving and those born and living for long periods (aboriginals) in the Sukhumi farm, was carried out. The rate of detection of antibody to HAV (anti-HAV) was shown to vary from 47% (Papio hamadryas) to 100% (Macaca arctoides and Macaca fascicularis). The level of infection with HAV varied among different groups of the same species: Macaca rhesus from 30% to 96%, Papio hamadryas from 0 to 82%. During a long-term observation period seroconversion to HAV was observed in monkeys arriving to the farm from natural habitats. In M. rhesus upon arrival the anti-HAV were detected in 7% and by the end of the observation period reached 100%, in green monkeys 28% and 92%, respectively. Anti-HAV of the IgM class were detected in animals with seroconversion. In fecal extracts from M. rhesus and in the liver, feces, and intestinal contents of green monkeys HAV was found cross-reacting with simian and human sera containing anti-HAV. The virions isolated from a green monkey liver had a buoyant density in CsCl 1.36 g/cm3, and HAV from feces of a M. rhesus sedimented in the density zone of 1.34 g/cm3.     

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced acute transient dystonia in monkeys associated with low striatal dopamine

Unilateral intracarotid infusion of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in baboons produces transient contralateral dystonia lasting 2-3 weeks followed by chronic hemiparkinsonism. We now extend this model to Macaca nemestrina and Macaca fascicularis. MPTP was infused unilaterally into the internal carotid artery of two M. nemestrina and 11 M. fascicularis. Effects were assessed with blinded clinical ratings of dystonia and Parkinsonism; [18F]-6-fluoro-DOPA (FDOPA) positron emission tomography; and postmortem measurements of striatal dopamine content. In two M. nemestrina, MPTP 0.4 mg/kg intracarotid produced acute dystonia within 24 h then chronic Parkinsonism starting 3 weeks later. In three M. fascicularis, MPTP 0.4 mg/kg produced acute dystonia within 3-8 h but two others died from large hemispheric infarcts within 1 day. A much lower dose, MPTP 0.1 mg/kg produced no clinical manifestations (n=1), whereas MPTP 0.25 mg/kg produced consistent transient dystonia and ipsiversive turning within 1-3 days followed by chronic Parkinsonism at 3 weeks (n=5). One week after MPTP, striatal FDOPA uptake decreased an average of 69% in M. nemestrina (0.4 mg/kg); and decreased an average of 70+/-21% in M. fascicularis (0.25 mg/kg). Striatal dopamine was reduced an average 66% in the first day (n=2) during acute dystonia, 98% at 3 days (n=1) and 99%+/-2.3% at 2-4 months (n=5). M. nemestrina had a clinical response similar to baboons whereas M. fascicularis seemed more sensitive to MPTP. These findings extend the model of MPTP-induced transient dystonia followed by chronic hemiparkinsonism to M. nemestrina and M. fascicularis and demonstrate that the early dystonic phase is accompanied by striatal dopamine deficiency.     

Lymphoid immunohistochemistry of macaque primates.

The immunohistochemical localizations of 53 monoclonal antibodies with specificities expressed in human lymphoid tissue were investigated on lymph node tissues from Macaca mulatta, M. speciosa, and M. fascicularis using immunoperoxidase techniques. Thirty antibodies showed comparative cross-reactivity. The micro-anatomic distribution of these antigens was closely homologous to that observed in man and antigen densities appeared to be similar. A useful monoclonal antibody panel for immunopathological investigations in macaques was, thereby, defined. Of several pan-T antibodies studied, only OKT11(CD2) showed cross-reactivity. A detailed analysis of the immunocyto-architecture of five rhesus lymph nodes was undertaken. Automated flow cytometry of M. mulatta and M. speciosa peripheral blood showed a broadly similar pattern to man.     

Susceptibility of Macaca rhesus to infection with hepatitis A virus strains isolated from man and monkeys

The results of infection of Macaca rhesus monkeys with hepatitis A virus (HAV) strains isolated from monkeys and man are presented. After inoculation of monkeys with human NAV strains the infection could not be reproduced whereas simian HAV strains were found to be pathogenic for M. rhesus monkeys both on experimental inoculation and during natural contacts with infected monkeys in the animal house.     

‘Long-loop’ reflexes can be obtained in spinal monkeys

Extensor muscles of the fore- and hindlimb were stretched in the lightly anaesthetized monkey (Macaca fascicularis) and cat. The resulting electromyogram contained a short latency peak consistent with a monosynaptic, segmental pathway; this peak was identified as the M1 peak of Tatton et al. [21]. A longer latency peak, identified as M2 and said to involve a pathway including supraspinal centres, was also present.After spinal section at a high cervical level, the electromyogram was reduced in amplitude, but both short (M1) and longer latency (M2) peaks were present. It is concluded that neither the cerebral cortex nor the cerebellum are necessary parts of the neural circuitry generating longer latency components of the electromyographic response to muscle stretch.     

Regulatory role of FcR+ and FcR- monocyte subsets in Mycobacterium leprae-induced lymphoproliferative response in vitro.

We investigated nine rhesus monkeys (Macaca mulatta) inoculated with Mycobacterium leprae and three normal human contacts. Peripheral blood monocytes were separated into Fc receptor positive (FcR+) and Fc receptor negative (FcR-) fractions, and their regulatory role in the lymphoproliferative response in vitro to M. leprae was studied. FcR- monocytes had strong antigen presentation activity and produced no suppressor effect while FcR+ monocytes had weak antigen presentation activity and produced a non-specific suppressor factor spontaneously. With this assay system we determined that M. leprae-inoculated rhesus monkeys could be divided into three groups: good responders, very weak responders, and non-responders.     

Variations in the structure of the prelunate gyrus in Old World monkeys

Anatomical and electrophysiological studies have revealed a complex organization in the macaque prelunate gyrus. We investigated the morphology and architecture of the prelunate gyrus in Old World monkeys. In Macaca nemestrina, we observed a sulcus crossing the prelunate gyrus within 2 mm of the vertical meridian representation. In other macaque species and other cercopithecines, we observed substantial variations in sulcal morphology across the prelunate gyrus. We did not find a sulcus in all species, and the location and depth of that indentation on the gyrus varied among species. A deep sulcus was observed in all species that emerged earlier in evolution than macaques, such as guenons, baboons, and colobines. We analyzed the regional and parcellation features of the prelunate gyrus in three macaque species, M. maura, M. mulatta, and M. radiata, and in Erythrocebus patas, with emphasis on the relation of structure to the distribution of prelunate visual areas. Nonphosphorylated neurofilament protein immunoreactivity permitted the delineation of a novel area in the prelunate gyrus of Old World monkeys, located around the prelunate sulcus. Species-specific patterns were also observed in the prelunate gyrus of the patas monkey compared to macaques. These observations, as well as a cladistic analysis of the data, suggest an expanded and diversified organization of the prelunate gyrus in some cercopithecoids that may reflect adaptation to specific ecological environments. It was, however, progressively lost in most macaques, being retained only in species that diverged early in the evolution of the genus Macaca, such as M. nemestrina and M. maura.     

Ultrastructural characteristics and lectin-binding properties of M cells in the follicle-associated epithelium of chicken caecal tonsils

To clarify the nature of M cells, the detailed ultrastructural characteristics and lectin-binding properties of M cells were investigated in follicle-associated epithelium (FAE) of chicken caecal tonsils. M cells presented various outlines from columnar to dome shaped. Their polymorphism was dependent on the number of harboured intraepithelial migrating cells. The lighter and larger nuclei of M cells were situated at more apical levels in the epithelial lining compared with those of neighbouring microvillous epithelial cells. The microvilli, which were significantly shorter and thicker than those of adjacent microvillous epithelial cells, were sparsely distributed or completely absent on the apical surfaces of M cells. In general, the apical cytoplasm of M cells without microvilli protruded slightly into the intestinal lumen. Numerous small vesicles were often contained in the apical cytoplasm. The numerous small invaginations of the apical and lateral cell surfaces suggested active transportation of luminal substances. No canaliculi existed in the apical cytoplasm of M cells whereas they were often detected in the neighbouring microvillous epithelial cells. A noteworthy finding was the frequent detection of multivesicular bodies in the apical cytoplasm of M cells. These multivesicular bodies suggest some degradation of ingested luminal substances during transcytoplasmic transportation. WGA and 4 other lectins strongly reacted with all epithelial cells except for M cells, this negativity suggesting a means of detecting M cells in chicken caecal tonsils. Three lectins, DSL, ConA and Jacalin, reacted weakly with the glycocalyx on M cells. The positive reactivity might allow chicken M cells to be utilised for specific antigen delivery into the mucosal immune system in some parenteral vaccinations.     

Allelic diversity within the high frequency Mamu-A2*05/Mane-A2*05 (Mane-A*06)/Mafa-A2*05 family of macaque MHC-A loci

Macaque species serve as important animal models of human infection and immunity. To more fully scrutinize their potential in both the analysis of disease pathogenesis and vaccine development, it is necessary to characterize the major histocompatibility complex (MHC) class I loci of Macaca mulatta (Mamu), Macaca nemestrina (Mane), and Macaca fascicularis (Mafa) at the genomic level. The oligomorphic Mamu-A2*05/Mane-A2*05 (previously known as Mane-A*06) family of macaque MHC-A alleles has recently been shown to be present at high frequency in both Indian rhesus and pig-tailed macaque populations. Using a locus-specific amplification and direct DNA typing methodology, we have additionally found that the locus encoding this family is very prevalent (75%) among a sampling of 182 Chinese rhesus macaques and has a high prevalence (80%) within a larger, independent cohort of 309 pig-tailed macaques. Interestingly, among the Chinese rhesus macaques, only six alleles previously identified in Indian-origin animals were observed, while three recently identified in Chinese-origin animals and 25 new alleles were characterized. Among the pig-tailed macaques, we observed 1 previously known (Mane-A*06) and 19 new alleles. Examination of the orthologous locus in a preliminary sampling of 30 cynomolgus macaques showed an even higher presence (87%) of Mafa-A2*05 family alleles, with 5 previously identified and 15 new alleles characterized. The continued discovery of novel alleles and thus further diversity within the Mamu-A2*05/Mane-A2*05/Mafa-A2*05 family indicates that this MHC-A locus, although highly conserved across the three species of macaques, has remained a dynamic entity during evolution.     

Local immune response to repeated topical antigen application in the simian labial mucosa.

Minor salivary glands of the oral mucosa in healthy monkeys (Macaca fascicularis and Macaca mulatta) contain organized structural units suitable for recognizing and processing antigens. A previous study of M. fascicularis monkeys provided experimental evidence of retrograde access of oral antigens deep into the minor salivary glands. The present study aimed at exploring the possible immune response of simian oral mucosa to repeated topical application of a chemically defined antigenic solution at the labial and gut mucosa. Ten female M. fascicularis animals were challenged topically at the lower lip mucosa at weekly intervals for a variable period of 4 to 8 weeks with a solution consisting of horseradish peroxidase, ferritin, and special India ink. Transmission electron microscopic examination of immunohistochemically treated sections of the labial glands revealed the presence of plasma cells containing specific anti-horseradish peroxidase antibody. These cells resided in the interacinar regions. Enteric and gut priming with the same antigen in four other monkeys, bypassing the oral mucosa, failed to reveal the presence of horseradish peroxidase-positive plasma cells in the labial mucosa of any of the four animals, although in one animal such cells could be identified in a mesenteric lymph node. This is suggestive of the existence, at least in primates, of a local immune response of the oral mucosa independent of systemic involvement.     

Cytological examination and cellular composition of bone marrow in healthy, adult, cynomolgus monkeys (Macaca fascicularis)

The purpose of this study was to evaluate the cellular composition of the bone marrow of cynomolgus monkeys (Macaca fascicularis). Femoral bone marrow smears from 23 healthy, adult animals (11 males and 12 females) were examined. For each animal, three femoral bone marrow smears were prepared immediately after euthanasia and stained with May-Grünwald-Giemsa. On two of the three smears available, and for each of these smears, a 500-cell differential count was performed and the myeloid: erythroid (M:E) ratio established. The M:E ratio for males varied from 0.67∶1.00 to 1.85∶1.00 with a mean of 1.03∶1.00 and for females from 0.67∶1.00 to 1.63∶1.00 with a mean of 1.02∶1.00. The mean percentage of granulocytic, lymphocytic, plasmacytic and erythroid series was 47.60, 5.44, 1.45 and 46.05% for males and 47.28, 5.12, 1.49 and 46.28% for females. No significant differences were noted between males and females. All cell lines were well represented and showed normal maturation in both sexes. Megakaryocytes were adequate in number and morphology in all animals. Cynomolgus monkeys showed a bone marrow composition similar to rhesus monkeys (Macaca mulatta). Cytological examination of bone marrow was found to be a simple and rapid procedure, well suited to the toxicological research environment. It provided excellent information on cell distribution, morphology and maturation of the haematopoietic system.     

太行山猕猴掌面肤纹嵴线的走向

目的 探讨太行山猕猴掌面肤纹嵴线走向特征。 方法 对52例（♀37例、♂15例）太行山猕猴(Macaca mulatta)掌面肤纹的嵴线走向进行调查,建立太行山猕猴掌面肤纹嵴线走向模型,并与日本猕猴进行比较。 结果 在种群水平上,太行山猕猴掌面指间Ⅱ区嵴线走向末端主要至2、5、6区, 6区频率最高;掌面指间Ⅲ区嵴线走向末端主要结束于6区,掌面小鱼际角1区域嵴线多呈现12/4/2走向;掌面肤纹嵴线走向侧别之间差异没有显著性（P>0.05）,为对称结构;雌雄性之间,掌面角1的嵴线走向差异具有显著性（P<0.05）,角7、角8嵴线走向性差不明显（P>0.05）;太行山猕猴和两组日本猕猴（Macaca fuscata）掌面肤纹嵴线走向相互之间种属差异明显（P<0.01）。 结论 猕猴肤纹的嵴线走向特征可以作为研究种属差异的一个指标。     

Cytological examination and cellular composition of bone marrow in healthy, adult, cynomolgus monkeys (Macaca fascicularis)

The purpose of this study was to evaluate the cellular composition of the bone marrow of cynomolgus monkeys (Macaca fascicularis). Femoral bone marrow smears from 23 healthy, adult animals (11 males and 12 females) were examined. For each animal, three femoral bone marrow smears were prepared immediately after euthanasia and stained with May-Grünwald-Giemsa. On two of the three smears available, and for each of these smears, a 500-cell differential count was performed and the myeloid: erythroid (M:E) ratio established. The M:E ratio for males varied from 0.67∶1.00 to 1.85∶1.00 with a mean of 1.03∶1.00 and for females from 0.67∶1.00 to 1.63∶1.00 with a mean of 1.02∶1.00. The mean percentage of granulocytic, lymphocytic, plasmacytic and erythroid series was 47.60, 5.44, 1.45 and 46.05% for males and 47.28, 5.12, 1.49 and 46.28% for females. No significant differences were noted between males and females. All cell lines were well represented and showed normal maturation in both sexes. Megakaryocytes were adequate in number and morphology in all animals. Cynomolgus monkeys showed a bone marrow composition similar to rhesus monkeys (Macaca mulatta). Cytological examination of bone marrow was found to be a simple and rapid procedure, well suited to the toxicological research environment. It provided excellent information on cell distribution, morphology and maturation of the haematopoietic system.     

PrimaTB STAT-PAK assay, a novel, rapid lateral-flow test for tuberculosis in nonhuman primates

Tuberculosis (TB) is the most important zoonotic bacterial disease in nonhuman primates (NHP). The current diagnostic method, the intradermal palpebral tuberculin test, has serious shortcomings. We characterized antibody responses in NHP against Mycobacterium tuberculosis to identify immunodominant antigens and develop a rapid serodiagnostic test for TB. A total of 422 NHP were evaluated, including 243 rhesus (Macaca mulatta), 46 cynomolgus (Macaca fascicularis), and 133 African green (Cercopithecus aethiops sabaeus) monkeys at five collaborative centers. Of those, 50 monkeys of the three species were experimentally inoculated with M. tuberculosis. Antibody responses were monitored every 2 to 4 weeks for up to 8 months postinfection by MultiAntigen Print ImmunoAssay with a panel of 12 recombinant antigens. All of the infected monkeys produced antibodies at various levels and with different antigen recognition patterns. ESAT-6 and MPB83 were the most frequently recognized proteins during infection. A combination of selected antigens which detected antibodies in all of the infected monkeys was designed to develop the PrimaTB STAT-PAK assay by lateral-flow technology. Serological evaluation demonstrated high diagnostic sensitivity (90%) and specificity (99%). The highest rate of TB detection was achieved when the skin test was combined with the PrimaTB STAT-PAK kit. This novel immunoassay provides a simple, rapid, and accurate test for TB in NHP.     

太行山猕猴下颌骨的异速生长分析

目的：探讨太行山猕猴下颌骨随年龄变化的形态学改变,为太行山猕猴的体质学、医学实验、进一步确定其分类地位提供可靠的依据。方法：30例太行山猕猴下颌骨标本,按其齿序及牙齿磨损程度的不同分成6组,并对其18项变量进行了测量、统计和分析。结果：下颌骨的有关变量与齿序和牙齿磨损程度成异速生长,即有关变量在生长、发育过程中的变化与其年龄相关,并且不同变量在不同年龄段呈现不同的生长模式。结论：下颌体各变量的变化趋势与其功能是相适应的,与猕猴的食物组成和其取食习性亦具有相关关系。     

Purification of zymogen granules from monkey parotid glands.

A method giving highly purified zymogen granules from Macaca irus and Cercopithecus aethiops parotid glands in reported. A 0.3 M sucrose medium for homogenization was supplemented with 10 mM tris/HCl, pH 7.3, and 0.1 mM lauric acid to stabilize the fragile monkey zymogen granules. Nuclei and cell debris were sedimented at 150 times g. A "crude" zymogen granule fraction was trapped in the 1.0 M sucrose layer of a discontinuous sucrose gradient at 1000 times g. Equilibrium centrifugation in a continuous sucrose gradient gave a fraction of zymogen granules at 1.85 M sucrose. Compared to the homogenate, this fraction exhibited about two-fold increase in zymogen granule marker, whereas mitochondrial marker was reduced to 1/4 and lysosomal marker to 1/2. A low level of contamination from other cell organelles was confirmed by electron microscopic investigation.     

Hepatitis E virus antibodies in the macaques and in the personnel serving the macaques of the Adler apery

There is evidence for the rather high detection rate of antibodies against hepatic E virus (HEV) was rather high in the macaques from the Adler apery in 1999-2005. Anti-HEV was detected in 232 (57.3%) out of 405 examined rhesus macaques (Macaca mulata) and in 16 (16%) out of 100 Java ones (M. fascicularis). The detection rate of anti-HEV ranged from 12.5 to 89.5%% among the rhesus macaques and from 5.9 to 37.5% among the Java ones. Class M anti-HEV was found only in 3 (4.3%) out of 69 Java macaques and in none of the rhesus ones. Of importance are the data of detection of anti-HEV in 3 (7.5%) out of 40 examined employees of the Research Institute of Medical Primatology, all 3 (18.8%) out of 10 employees looking after the monkeys that belonged to the highest-risk group. The epidemiological and epizootological aspects of this infection require further studies.