Progress in subclinical research of kidney xenotransplantation / 器官移植

Xenotransplantation is an efficient pathway to solve the problem of transplant organ source deficiency in clinical settings. With the increasing progress of gene editing technique and immune suppression regimen, important development has been achieved on researches regarding pig to non-human primate kidney xenotransplantation, which provides a good condition for the introduction of the technique in the clinical application. In view of the substantial difference between human and non-human primate, and to meet the needs of current ethic requirements, it is necessary to perform subclinical studies for pig to human kidney xenotransplantation. In recent years, such subclinical studies with regard to the genetically modified pig to brain death recipient kidney xenotransplantation had been performed, indicating that kidney xenotransplantation gradually began to transit to the clinical development stage. However, donor/recipient selection and immune suppression regimen has not reached a consensus yet, and has to be clarified in subclinical studies. In this article, the current status and confronted problems of donor/recipient selection, immune suppression regimen and post transplantation management in the subclinical studies of kidney xenotransplantation were reviewed, aiming to promote the clinical transformation of kidney xenotransplantation to the clinical application.

Present situation and progress of xenotransplantation at home and abroad / 器官移植

Organ shortage has become one of the major challenges hindering the development of organ transplantation. Xenotransplantation is one of the most valuable methods to resolve global organ shortage. In recent years, the development of genetic engineering technique and research and development of new immunosuppressant have provided novel theoretical basis for xenotransplantation. International scholars have successively carried out researches on xenotransplantation in genetically modified pigs to non-human primates or brain death recipients, making certain substantial progresses. However, most of the researches are still in the preclinical stage, far from clinical application. Therefore, according to the latest preclinical experimental research progress at home and abroad, the history of xenotransplantation, the development of gene modification technology, xenotransplantation rejection and immunosuppression regimens were reviewed, aiming to provide reference for subsequent research of xenotransplantation, promote clinical application of xenotransplantation and bring benefits to more patients with end-stage diseases.

Application of xenotransplantation in clinical practice / 器官移植

Organ transplantation is the optimal treatment for end-stage organ failure. Nevertheless, organ shortage is a global problem, which limits further development of organ transplantation. Recent research shows that genetically modified pig may become a realistic alternative source of clinical organ transplantation donor. Xenotransplantation may serve as one of the effective measures to resolve the problem of organ shortage. Since 2021, 2 cases of living xenotransplantation and 6 cases of xenotransplantation in brain death recipients have been performed worldwide, and phase Ⅰ clinical trial of xenotransplantation has been launched, and the results have exceeded expectations. Therefore, in this article, recent clinical trial results of xenotransplantation in living and brain death recipients were retrospectively analyzed, and scientific, technical and ethical issues related to clinical research of xenotransplantation were illustrated, hoping to provide reference for clinical research of xenotransplantation in China and promote the development of xenotransplantation in clinical practice.

A non-human primate derived anti-P-selectin glycoprotein ligand-1 antibody curtails acute pancreatitis by alleviating the inflammatory responses

Acute pancreatitis (AP) is a devastating disease characterized by an inflammatory disorder of the pancreas. P-selectin glycoprotein ligand-1 (PSGL-1) plays a crucial role in the initial steps of the adhesive at process to inflammatory sites, blockade of PSGL-1 might confer potent anti-inflammatory effects. In this study, we generated two non-human primate derived monoclonal antibodies capable of efficiently targeting human PSGL-1, RH001-6 and RH001-22, which were screened from immunized rhesus macaques. We found that RH001-6, can effectively block the binding of P-selectin to PSGL-1, and abolish the adhesion of leukocytes to endothelial cells in vitro. In vivo, we verified that RH001-6 relieved inflammatory responses and pancreatic injury in both caerulein and l-arginine induced AP models. We also evaluated the safety profile after RH001-6 treatment in mice, and verified that RH001-6 did not cause any significant pathological damages in vivo. Taken together, we developed a novel non-human primate derived PSGL-1 blocking antibody with high-specificity, named RH001-6, which can interrupt the binding of PSGL-1 and P-selectin and attenuate inflammatory responses during AP. Therefore, RH001-6 is highly potential to be further developed into therapeutics against acute inflammatory diseases, such as AP.

Genetic Approaches for Neural Circuits Dissection in Non-human Primates / 神经科学通报·英文版

Genetic tools, which can be used for the morphology study of specific neurons, pathway-selective connectome mapping, neuronal activity monitoring, and manipulation with a spatiotemporal resolution, have been widely applied to the understanding of complex neural circuit formation, interactions, and functions in rodents. Recently, similar genetic approaches have been tried in non-human primates (NHPs) in neuroscience studies for dissecting the neural circuits involved in sophisticated behaviors and clinical brain disorders, although they are still very preliminary. In this review, we introduce the progress made in the development and application of genetic tools for brain studies on NHPs. We also discuss the advantages and limitations of each approach and provide a perspective for using genetic tools to study the neural circuits of NHPs.

Kidney xenotransplantation: status quo and development trend of physiological research / 器官移植

Organ transplantation is the most effective treatment for all categories of end-stage organ diseases. To resolve the shortage of donors in organ transplantation, widespread attention has been diverted to xenotransplantation. At present, clinicians mainly highlight the problems related to xenotransplantation rejection and viral infection. The physiology of xenotransplantation has been rarely studied. Kidney performs endocrine function by producing erythropoietin (EPO), renin and activating vitamin D. Although these pathways are usually well preserved in allogeneic transplantation, species-specific differences, especially those between pigs and non-human primates, may still affect the physiological function of transplant organs. In this article, the changes of EPO, renin-angiotensin-aldosterone system (RAAS) and active vitamin D3 of pig and human after xenotransplantation were illustrated, aiming to provide reference for subclinical research of xenotransplantation.

Fast surveillance response reveals the introduction of a new yellow fever virus sub-lineage in 2021, in Minas Gerais, Brazil

BACKGROUND In Brazil, the yellow fever virus (YFV) is maintained in a sylvatic cycle involving wild mosquitoes and non-human primates (NHPs). The virus is endemic to the Amazon region; however, waves of epidemic expansion reaching other Brazilian states sporadically occur, eventually causing spillovers to humans. OBJECTIVES To report a surveillance effort that led to the first confirmation of YFV in NHPs in the state of Minas Gerais (MG), Southeast region, in 2021. METHODS A surveillance network was created, encompassing the technology of smartphone applications and coordinated actions of several research institutions and health services to monitor and investigate NHP epizootics. FINDINGS When alerts were spread through the network, samples from NHPs were collected and YFV infection confirmed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and genome sequencing at an interval of only 10 days. Near-complete genomes were generated using the Nanopore MinION sequencer. Phylogenetic analysis indicated that viral genomes were related to the South American genotype I, clustering with a genome detected in the Amazon region (state of Pará) in 2017, named YFVPA/MG sub-lineage. Fast YFV confirmation potentialised vaccination campaigns. MAIN CONCLUSIONS A new YFV introduction was detected in MG 6 years after the beginning of the major outbreak reported in the state (2015-2018). The YFV strain was not related to the sub-lineages previously reported in MG. No human cases have been reported, suggesting the importance of coordinated surveillance of NHPs using available technologies and supporting laboratories to ensure a quick response and implementation of contingency measures to avoid YFV spillover to humans.

Generation of a Hutchinson-Gilford progeria syndrome monkey model by base editing

Many human genetic diseases, including Hutchinson-Gilford progeria syndrome (HGPS), are caused by single point mutations. HGPS is a rare disorder that causes premature aging and is usually caused by a de novo point mutation in the LMNA gene. Base editors (BEs) composed of a cytidine deaminase fused to CRISPR/Cas9 nickase are highly efficient at inducing C to T base conversions in a programmable manner and can be used to generate animal disease models with single amino-acid substitutions. Here, we generated the first HGPS monkey model by delivering a BE mRNA and guide RNA (gRNA) targeting the LMNA gene via microinjection into monkey zygotes. Five out of six newborn monkeys carried the mutation specifically at the target site. HGPS monkeys expressed the toxic form of lamin A, progerin, and recapitulated the typical HGPS phenotypes including growth retardation, bone alterations, and vascular abnormalities. Thus, this monkey model genetically and clinically mimics HGPS in humans, demonstrating that the BE system can efficiently and accurately generate patient-specific disease models in non-human primates.

Acute effects of ayahuasca in a juvenile non-human primate model of depression

Objective: The incidence rate of major depression in adolescents reaches approximately 14%. This disorder is usually recurrent, without remission of symptoms even after pharmacological treatment, and persists throughout adult life. Since the effects of antidepressants take approximately 2 weeks to begin, new pharmacological therapies are under continuous exploration. Recent evidence suggests that psychedelics could produce rapid antidepressant effects. In this study, we evaluated the potential antidepressant effects of ayahuasca in a juvenile non-human primate model of depression. Methods: While living with their families, juvenile marmosets (8 males; 7 females) were observed on alternate days for four weeks during a baseline phase. This was followed by 8 weeks of an induced depressive state protocol, the social isolated context (IC), in which the animals were monitored in the first and last weeks. Subsequently, five males and four females were randomly selected for treatment, first with a single administration of saline vehicle (1.67 mL/300 g of body weight, via gavage), followed by a single dose of ayahuasca (1.67 mL/300 g of body weight, via gavage). Both phases lasted 1 week and the animals were monitored daily. A third week of sampling was called the tardive-pharmacological effects phase. In all phases the marmosets were assessed for behavior, fecal cortisol levels, and body weight. Results: After IC, the animals presented typical hypocortisolemia, but cortisol recovered to baseline levels 24 h after an acute dose of ayahuasca; this recovery was not observed in vehicle-treated animals. Additionally, in males, ayahuasca, but not the vehicle, reduced scratching, a stereotypic behavior, and increased feeding. Ayahuasca treatment also improved body weight to baseline levels in both sexes. The ayahuasca-induced behavioral response had long-term effects (14 days). Thus, in this translational juvenile animal model of depression, ayahuasca presented beneficial effects. Conclusions: These results can contribute to the validation of ayahuasca as an antidepressant drug and encourage new studies on psychedelic drugs as a tool for treating mood disorders, including for adolescents with early-onset depression.

Molecular identification of hemoplasmas in free ranging non-human primates in Thailand

Objective: To survey hemoplasmas infection in free ranging non-human primates from 8 provinces in Thailand. Methods: DNA from ethylenediaminetetraacetic acid blood of 262 free ranging non-human primates were identified as hemoplasmas using PCR and phylogenetic analysis based on 16S rRNA and rnpB genes. Results: A total of 148 non-human primates (56.49%) were determined positive for Candidatus Mycoplasma haemomacaque, including 125 Macaca fascicularis and 23 Macaca mulatta. Hemoplasmas can cause anemia in monkey but all positive samples were healthy. The positive rates in male and female non-human primates were not significantly different. Conclusions: Candidatus Mycoplasma infection is prevalent in free ranging Macaca fascicularis and Macaca mulatta in Thailand.

Establishment of acute liver failure models induced by D-galactosamine in non-human primates / 器官移植

Objective To evaluate the effect of different doses of D-galactosamine (D-Gal) on non-human primate cynomolgus monkey and to establish a monkey model with different degree of acute liver failure (ALF). Methods Twelve cynomolgus monkeys were evenly divided into the low-, medium- and high-dose groups (n=4) with a dosage of 0.23, 0.25 and 0.27 g/kg, respectively. In each group, the corresponding dose of D-Gal solution was injected into the monkeys through the forearm vein at one time in a sober state (without anesthesia). The survival time of the cynomolgus monkeys was recorded. Digestive tract and hepatic encephalopathy symptoms were observed. Vital signs were measured at 0 h before and 12, 24, 36, 48, 60, 72, 96, 120 and 144 h after D-Gal administration. Alanine transaminase (ALT), total bilirubin (TB), prothrombin time (PT), blood ammonia and other parameters were detected from the blood samples. The liver tissues were prepared for hematoxylin-eosin (HE) staining to observe the pathological changes. Results All cynomolgus monkeys in the low-dose group survived and transient liver injury was noted without the hepatic encephalopathy symptoms. At 60 h after D-Gal administration, the liver function and coagulation indexes reached the peak, gradually recovered and then basically returned to the normal range at 120 h. In the medium-dose group, the course of disease was relatively slow and gradually recovered after the appearance of severe liver damage and hepatic encephalopathy symptoms and only one animal died. All cynomolgus monkeys in the high-dose group died after developing hepatic encephalopathy symptoms and severe liver damage with a mean survival time of (72±13) h. Pathological examination of liver tissue demonstrated that scattered liver cell necrosis and inflammatory cell infiltration were observed in the liver tissues of the low-dose group. In the medium- and high-dose groups, the hepatic lobule structure was not clear, and the liver cell necrosis in flakes accompanied by evident hemorrhage were documented. Conclusions The D-Gal dosage in the medium- and high-dose groups meet the standards of the ALF model. The degree of ALF in the medium-dose group is relatively slight, which is beneficial to the implementation of liver transplantation. ALF in the high-dose group is relatively severe, which is suitable for the evaluation of the clinical efficacy of therapeutic options.

Molecular identification of hemoplasmas in free ranging non-human primates in Thailand

Objective: To survey hemoplasmas infection in free ranging non-human primates from 8 provinces in Thailand. Methods: DNA from ethylenediaminetetraacetic acid blood of 262 free ranging non-human primates were identified as hemoplasmas using PCR and phylogenetic analysis based on 16S rRNA and rnpB genes. Results: A total of 148 non-human primates (56.49%) were determined positive for Candidatus Mycoplasma haemomacaque, including 125 Macaca fascicularis and 23 Macaca mulatta. Hemoplasmas can cause anemia in monkey but all positive samples were healthy. The positive rates in male and female non-human primates were not significantly different. Conclusions: Candidatus Mycoplasma infection is prevalent in free ranging Macaca fascicularis and Macaca mulatta in Thailand.

Abnormal Mitochondria in a Non-human Primate Model of MPTP-induced Parkinson's Disease: Drp1 and CDK5/p25 Signaling

Mitochondria continuously fuse and divide to maintain homeostasis. An impairment in the balance between the fusion and fission processes can trigger mitochondrial dysfunction. Accumulating evidence suggests that mitochondrial dysfunction is related to neurodegenerative diseases such as Parkinson's disease (PD), with excessive mitochondrial fission in dopaminergic neurons being one of the pathological mechanisms of PD. Here, we investigated the balance between mitochondrial fusion and fission in the substantia nigra of a non-human primate model of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD. We found that MPTP induced shorter and abnormally distributed mitochondria. This phenomenon was accompanied by the activation of dynamin-related protein 1 (Drp1), a mitochondrial fission protein, through increased phosphorylation at S616. Thereafter, we assessed for activation of the components of the cyclin-dependent kinase 5 (CDK5) and extracellular signal-regulated kinase (ERK) signaling cascades, which are known regulators of Drp1(S616) phosphorylation. MPTP induced an increase in p25 and p35, which are required for CDK5 activation. Together, these findings suggest that the phosphorylation of Drp1(S616) by CDK5 is involved in mitochondrial fission in the substantia nigra of a non-human primate model of MPTP-induced PD.

Nephroblastoma in a black-tufted marmoset (Callithrix penicillata) / Nefroblastoma em um sagui-de-tufos-pretos (Callithrix penicillata)

A renal nephroblastoma is described in a free-living black-tufted marmoset (Callithrix penicillata) in Central Brazil. The monkey was found dead and subjected to necropsy. Gross anatomic changes consisted of a ruptured left kidney, which was almost completely effaced by a white to yellow, partially encapsulated friable mass. The left ureter was distended due to obstruction by a red, spherical, 2mm in diameter friable mass. The urinary bladder was also distended. Histologically the renal and ureteral masses consisted of a triphasic embryonal neoplasm composed of embryonic epithelium forming glomeruli and tubules, polygonal blastemal cells, and a mesenchymal stroma. The embryonic epithelium exhibited rare nuclear immunoreactivity for WT-1, whereas blastemal cells exhibited robust cytoplasmic and rare nuclear immunoreactivity for WT-1; blastemal cells were also immunoreactive for vimentin. No immunoreactivity was detected for pan-cytokeratin (AE1/AE3), actin, and desmin. Morphological and immunohistochemical features of the present neoplasm are consistent with those described for renal nephroblastoma.(AU)

Descreve-se um caso de nefroblastoma maligno em um sagui de vida livre no Brasil Central. O macaco foi encontrado morto e encaminhado para necropsia. Na macroscopia, o rim esquerdo apresentava-se rompido e o parênquima estava substituído por um tecido neoplásico friável, parcialmente encapsulado e de superfície natural branca e de corte amarela. O ureter esquerdo apresentava-se distendido devido à obstrução por uma massa friável, vermelha, esférica, de 2mm de diâmetro. Histologicamente, as massas renal e ureteral consistiam de uma neoplasia embrionária composta por três populaçõies de células neoplásicas, composta por epitélio embrionário formando glomérulos e túbulos, células blastemais poligonais e um estroma mesenquimal. O epitélio embrionário exibiu imunorreactividade nuclear rara para WT-1, enquanto que as células blastemais exibiram imunorreactividade nuclear citoplasmática e rara para WT-1; As células blastemais também foram imunorreativas à vimentina. Nenhuma imunorreatividade foi detectada para pan-citoqueratina (AE1/AE3), actina e desmina. As características morfológicas e imuno-histoquímicas da presente neoplasia são consistentes com as descritas para o nefroblastoma renal.(AU)

Pelvimetry in Squirrel Monkeys (Saimiri scireus Linnaeus, 1758) / Pelvimetria em macacos-de-cheiro (Saimiri scireus Linnaeus, 1758)

Saimiri sciureus is a neotropical primate widely used in research. However, there are still difficulties regarding their reproduction in vivaria due to the high incidence of dystocia. Dystocia in primates can be caused by cephalopelvic disproportion and in Squirrel Monkeys, pregnancy of large fetuses were reported. This paper describes pelvimetry data of adult females and subadults in specimens of Squirrel Monkeys, from a research colony of Para, Brazil. Pelvic radiographs were obtained in ventrodorsal projections and the following measurements were taken: superior bi-iliac diameter (SBID); inferior bi-iliac diameter (IBID); bi-iliac average diameter (BIAD); right diagonal diameter (RDD); left diagonal diameter (LDD); sacro-pubic diameter (SPD); Based on the obtained diameters, the entrance area of the pelvis (EAP) was also calculated. The average values of the pelvic diameters and EAP in adult females were SBID 1.714cm, BIAD 1.957cm, IBID 1.686cm, RDD 2.771cm, LDD 2.764cm, SPD 2.543cm and EAP 3.9056cm2; and subadult females: 1.588cm SBID, 1.850cm BIAD, 1.625cm IBID, 2.50cm RDD, LDD 2.474cm, 1.95cm SPD and 2.8293 cm2 EAP. Saimiri sciureus pelvis is characterized as dolichopelvic. There was statistical significance between the values for adult females and subadults to SBID, BIAD, RDD, LDD, SPD and EAP. The values of SBID and IBID were lower when compared to the published data for the same species. The result found on this paper will serve as a basis for future studies using pelvic measurements and dystocia prediction of neotropical primates and comparison between different vivaria.(AU)

Saimiri sciureus é uma espécie de primata neotropical muito utilizada como animal de pesquisa. No entanto ainda há dificuldades em biotérios quanto a sua reprodução devido à alta ocorrência de distocia. A distocia em primatas pode ter origem devido à desproporção cefalopélvica, sendo que em macacos-de-cheiro é relatada a gestação de fetos grandes. O presente trabalho descreve dados de pelvimetria em espécimes de macaco-de-cheiro, fêmeas adultas e subadultas provenientes de uma colônia de pesquisa do Pará, Brasil. Foram realizadas radiografias da pelve em projeção ventrodorsal e por meio destas mensurados os diâmetros biilíaco superior (DBIS); diâmetro biíliaco inferior (DBII); diâmetro biilíaco médio (DBIM); diâmetro diagonal direito (DDD); diâmetro diagonal esquerdo (DDE); diâmetro sacro-púbico (DSP); com base nos diâmetros obtidos também foi calculada a área de entrada da pelve (AEP). Os valores médios dos diâmetros pélvicos e da AEP em fêmeas adultas foram: DBIS 1,714cm, DBIM 1,957cm, DBII 1,686cm, DDD 2,771cm, DDE 2,764cm, DSP 2,543cm e AEP 3,9056cm2; e para fêmeas subadultas: 1,588cm DBIS, 1,850cm DBIM, 1,625cm DBII, 2,50cm DDD, 2,474cm DDE, 1,95cm DSP e 2,8293 cm2 AEP. Observou-se que a pelve de Saimiri sciureus é dolicopélvica. Houve diferença estatística significativa entre os valores para fêmeas adultas e subadultas para DBIS, DBIM, DDD, DDE, DSP e AEP. Em comparação com dados da literatura de pelvimetria para S. sciureus observaram-se menores valores de DBIS e DBII. O resultado deste trabalho servirá como base para futuros estudos utilizando-se mensurações pélvicas e predição de distocia em primatas neotropicais e referência para comparação entre S. sciureus de diferentes biotérios.(AU)

Cortical Representation of Pain and Touch: Evidence from Combined Functional Neuroimaging and Electrophysiology in Non-human Primates / 神经科学通报·英文版

Human functional MRI studies in acute and various chronic pain conditions have revolutionized how we view pain, and have led to a new theory that complex multi-dimensional pain experience (sensory-discriminative, affective/motivational, and cognitive) is represented by concurrent activity in widely-distributed brain regions (termed a network or pain matrix). Despite these breakthrough discoveries, the specific functions proposed for these regions remain elusive, because detailed electrophysiological characterizations of these regions in the primate brain are lacking. To fill in this knowledge gap, we have studied the cortical areas around the central and lateral sulci of the non-human primate brain with combined submillimeter resolution functional imaging (optical imaging and fMRI) and intracranial electrophysiological recording. In this mini-review, I summarize and present data showing that the cortical circuitry engaged in nociceptive processing is much more complex than previously recognized. Electrophysiological evidence supports the engagement of a distinct nociceptive-processing network within SI (i.e., areas 3a, 3b, 1 and 2), SII, and other areas along the lateral sulcus. Deafferentation caused by spinal cord injury profoundly alters the relationships between fMRI and electrophysiological signals. This finding has significant implications for using fMRI to study chronic pain conditions involving deafferentation in humans.

Decoding Saccadic Directions Using Epidural ECoG in Non-Human Primates

A brain-computer interface (BCI) can be used to restore some communication as an alternative interface for patients suffering from locked-in syndrome. However, most BCI systems are based on SSVEP, P300, or motor imagery, and a diversity of BCI protocols would be needed for various types of patients. In this paper, we trained the choice saccade (CS) task in 2 non-human primate monkeys and recorded the brain signal using an epidural electrocorticogram (eECoG) to predict eye movement direction. We successfully predicted the direction of the upcoming eye movement using a support vector machine (SVM) with the brain signals after the directional cue onset and before the saccade execution. The mean accuracies were 80% for 2 directions and 43% for 4 directions. We also quantified the spatial-spectro-temporal contribution ratio using SVM recursive feature elimination (RFE). The channels over the frontal eye field (FEF), supplementary eye field (SEF), and superior parietal lobule (SPL) area were dominantly used for classification. The α-band in the spectral domain and the time bins just after the directional cue onset and just before the saccadic execution were mainly useful for prediction. A saccade based BCI paradigm can be projected in the 2D space, and will hopefully provide an intuitive and convenient communication platform for users.

Advance of gene-modified non-human primate models / 生物工程学报

Non-human primates would be particularly valuable in life sciences and biomedical research area. Gene-modified monkeys with gene overexpression or loss of function have been successfully generated with the rapid advance in gene manipulation technology such as lentivirus infection and programmable nucleases (ZFN, TALEN, CRISPR-Cas9). Here we review the recent development on gene-modified monkey generation by lentivirus and programmable nucleases. Then we discuss three concerns, the long time for sexual maturation, the off target and the mosaicism of founders, which limit the wide application of gene-modified non-human-primates. At last, hotspots and future trend for gene-modified non-human-primates generation are proposed.

Increase in Anti-Gal IgM Level is Associated With Early Graft Failure in Intraportal Porcine Islet Xenotransplantation

BACKGROUND: Anti-Gal is a major antibody induced in non-human primates (NHPs) after xenotransplantation. To understand the mechanism of graft rejection, we investigated the association between anti-Gal responses and graft failure in NHP recipients of porcine islet transplantation (PITx). METHODS: Intraportal PITx was performed in 35 diabetic NHPs, and graft function was monitored. Early graft failure (EGF) was defined as loss of graft function within a month after PITx. Seven, 19, nine NHPs received immunosuppression (IS) without CD40 pathway blockade (Group I), with anti-CD154 (Group II), and with anti-CD40 (Group III), respectively. The anti-Gal levels on day 0 and day 7 of PITx were measured by ELISA. RESULTS: The frequency of EGF was significantly lower in Group II (26.3%) than in Group I (100%, P=0.0012) and Group III (77.8%, P=0.0166). While levels of anti-Gal IgG in Group I and anti-Gal IgM in Group III increased on day 7 compared with day 0 (P=0.0156 and 0.0273), there was no increase in either on day 7 in Group II. The ratio of anti-Gal IgM or IgG level on day 7 to that on day 0 (Ratio7/0) was significantly higher in recipients with EGF than without EGF (P=0.0009 and 0.0027). ROC curve analysis of anti-Gal IgM Ratio7/0 revealed an area under the curve of 0.789 (P=0.0003). CONCLUSIONS: IS with anti-CD154 suppressed anti-Gal responses and prevented EGF in PITx. Anti-Gal IgM Ratio7/0, being associated with EGF, is a predictive marker for EGF.

Cryptosporidium spp., Giardia intestinalis, and Enterocytozoon bieneusi in Captive Non-Human Primates in Qinling Mountains

Non-human primates (NHPs) are confirmed as reservoirs of Cryptosporidium spp., Giardia intestinalis, and Enterocytozoon bieneusi. In this study, 197 fresh fecal samples from 8 NHP species in Qinling Mountains, northwestern China, were collected and examined using multilocus sequence typing (MLST) method. The results showed that 35 (17.8%) samples were positive for tested parasites, including Cryptosporidium spp. (3.0%), G. intestinalis (2.0%), and E. bieneusi (12.7%). Cryptosporidium spp. were detected in 6 fecal samples of Macaca mulatta, and were identified as C. parvum (n=1) and C. andersoni (n=5). Subtyping analysis showed Cryptosporidium spp. belonged to the C. andersoni MLST subtype (A4, A4, A4, and A1) and C. parvum 60 kDa glycoprotein (gp60) subtype IId A15G2R1. G. intestinalis assemblage E was detected in 3 M. mulatta and 1 Saimiri sciureus. Intra-variations were observed at the triose phosphate isomerase (tpi), beta giardin (bg), and glutamate dehydrogenase (gdh) loci, with 3, 1, and 2 new subtypes found in respective locus. E. bieneusi was found in Cercopithecus neglectus (25.0%), Papio hamadrayas (16.7%), M. mulatta (16.3%), S. sciureus (10%), and Rhinopithecus roxellana (9.5%), with 5 ribosomal internal transcribed spacer (ITS) genotypes: 2 known genotypes (D and BEB6) and 3 novel genotypes (MH, XH, and BSH). These findings indicated the presence of zoonotic potential of Cryptosporidium spp. and E. bieneusi in NHPs in Qinling Mountains. This is the first report of C. andersoni in NHPs. The present study provided basic information for control of cryptosporidiosis, giardiasis, and microsporidiosis in human and animals in this area.