Large granular lymphocytosis in a cynomolgus macaque (Macaca fascicularis) with a subclinical Trypanosoma cruzi infection

A 5.25-year-old cynomolgus macaque (Macaca fascicularis) was found to have a marked leukocytosis due to a lymphocytosis on routine quarantine laboratory data prior to inclusion in a preclinical research study. The majority of lymphocytes were characterized as intermediate to large with round to convoluted nuclei, coarse to clumped chromatin, rare prominent nucleoli, and moderate amounts of lightly basophilic cytoplasm that frequently contained small magenta granules and/or clear vacuoles. The animal had tested negative for several viruses and other etiologic agents found in nonhuman primates 1 week prior to shipment to the research facility. However, further evaluation of the blood smear revealed rare hemoflagellates, and later testing using real-time PCR and ELISA was confirmatory for Trypanosoma cruzi (T cruzi). Trypanosoma cruzi is a zoonotic pathogen responsible for Chagas disease in people and can have negative consequences on study results when positive animals are inadvertently used for preclinical research. This case report describes a marked large granular lymphocytosis in an otherwise healthy macaque as the only indication of infection with T cruzi in an animal believed to be negative for the infection. Additionally, it highlights the diagnostic limitations of screening tests to rule out diseases in animals intended to be used in preclinical studies.     

雏鸡感染传染性法氏囊病毒后血液的T细胞变化

本项研究对雏鸡感染传染性法氏囊病毒(IBDV)后,1～8周外周血液的 ANAE~+淋巴细胞(T 细胞)、淋巴细胞和白细胞数量的动态变化进行了检测。结果表明,1日龄雏鸡感染 IBDV 后,其血液中 ANAE~+,淋巴细胞数量明显低于对照雏鸡,表明其细胞免疫机能降低;4周龄雏鸡感染 IBDV 后,其 ANAE~+淋巴细胞数量呈一过性降低,变化不如1日龄感染雏鸡明显,说明 IBDV 对雏鸡存在感染年龄差异;对 ANAE~+淋巴细胞绝对数的检测较百分比能更真实地反映 IBD 鸡的细胞免疫功能变化的实质。     

饲料中添加花粉和酵母硒对中华鳖幼鳖生长和非特异性免疫功能的影响

在基础饲料中分别添加0.5×10-6酵母硒、0.1%花粉、0.5×10-6酵母硒+0.1%花粉的混合物,投喂中华鳖幼鳖90d后。测定幼鳖的生长和血清中谷胱甘肽过氧化物酶(GSH PX)、超氧化物歧化酶(SOD)活性、T淋巴细胞转化率、自然杀伤细胞(NK)活性。同对照组相比,0.5×10-6酵母硒+0.1%花粉组能显著促进中华鳖的生长(P<0.05),0.5×10-6酵母硒组与0.1%花粉组的日增重率均高于对照组,但差异不显著(P>0.05)。各组间成活率无差异(P>0.05)。0.5×10-6酵母硒组NK细胞活性以及血清中SOD酶和GSH PX酶的活力分别比对照组高了156.06%、44.7%、47.5%(P<0.05)。0.1%花粉组、0.5×10-6酵母硒+0.1%花粉组血清中GSH PX酶的活力分别比对照组高了39.9%、32.9%(P<0.05),但两组的SOD酶活力与对照组相比均无显著性差异(P>0.05)。0.5×10-6酵母硒组以及0.1%花粉组的T淋巴细胞转化率均比对照组高,但差异均不显著(P>0.05)。0.1%花粉组的NK细胞活性也比对照组高,但差异不显著(P>0.05)。0.5×10-6酵母硒+0.1%花粉组能显著增强中华鳖T淋巴细胞转化率及NK细胞活性(P<0.05)。     

Development of Immune Response to Experimental Bovine Trichophyton vervucosum Infection

Abstract— Cell mediated and humoral immune responses to experimental Trichophyton verrucosum infection were assessed by sequential cutaneous biopsies, antibody assessments and microscopic monitoring of fungal presence. Histopathologic examination showed the accrual of lymphocytes and other inflammatory cells in the dermis of infected sites. Immunoperoxidase staining of frozen sections with monoclonal antibody preparations revealed an influx of macrophages, BoCD4+ and B0CD8+ lymphocytes and γδ T cells from the 5th day to the 33rd day of infection. A moderate influx of B cells was observed. Protein G-colloidal gold staining revealed the presence of immunoglobulins in the dermis and superficial epidermal layers. Trichophyton specific serum antibodies appeared between days 33 and 55. Microscopic assessment of infected tissues revealed an increase in T, verrucosum elements (mycelium and ectothrix spores) from days 19 to 55. Fungal elements in infected areas did not decrease until after both humoral and cell mediated elements of the immune response were established. These responses imply a combination of cell mediated and humoral events were associated with T. verrucosum immunity and clearance in the calf. Résumé— Le réponse immunitaire humorale et cellulaire a une infection expérimentale àT. verrucosum a été appréciée par des biospsies cutanées successives, des dosages d'anticorps et la recherche microscopique de champignons. L'examen histopathologique a montré un afflux de lymphocytes et d'autres cellules inflammatoires dane le derme des sites infectés. Les marquages en immunopéroxydase par un anticorps monoclonal de coupes congelées a montré un influx de macrophages, lymphocytes BoCD4+ et BoCD8+ et des cellules T γδ, du 5e au 33e jour de l'infection. Un marquage par une protéine G—or colloidal a révélé d'immunogolglobulines dans le derme et les couches supéerficielles de l'épiderme. Les anticorps spécifiques de Trichophyton sont apparus entre 33 et 55 jours. L'examen microscopique des tissus infectés a révélé une augmentation du nombre d'éléments de T. verrucosum (mycelium et spores ectothrix) du 19e au 55e jours. Les éléments fongiques dans les zones infectées n'ont pas diminué avant que les réponses humorales et cellulaires ne solent établies. Ces résponses impliquent qu'une coopération des réponses humorales et cellulaires étaient associées dans l'immunité et la défense contre T. verrucosum. Zusammenfassung— Die zellvermittelten und humoralen Immunantworten auf die experimentelle Infektion mit T. verrucosum wurden durch eine Serie von Hautbiopsien, Antikörperuntersuchungen und mikroskopischer Untersuchung auf das Vorhandensein von Pilzen ausgewertet. Die histopathologische Untersuchung zeigte eine Ansammlung von Lymphozyten und anderen Entzündungszellen in der Dermis der infizierten Stellen. Die Immunperoxidasefärbung der Gefrierschnitte mit monoklonalen Antikörperzubereitungen zeigte einen Influx von Makrophagen, BoCd4-und BoCD8-Lymphozyten und gamma-delta-T-Zellen vom 5. bis zum 33. Tag der Infektion. Es wurde auch ein mäßiger Influx von B-Zellen beobachtet. Die Protein-G-kolloidale Goldfärbung zeigte die Anwesenheit von Immunglobulinen in der Dermis und den oberflächlichen epidermalen Schichten. Trichophyton-spezifische Serumantikörper traten zwischen Tag 33 und 55 auf. Die mikroskopische Untersuchung infizierter Gewebe zeigte eine Zunahme von T. verrucosum-Bestandteilen (Myzel und exktothrixe Sporen) vom Tag 19 bis 55. Pilzteile in infizierten Bereichen verminderten sich weder, nachdem humorale, noch nachdem zellvermittelte Elemente der Immunantwort auftraten. Diese Reaktionen deuten an, daß eine Kombination von zellvermittelten und humoralen Vorgängen im Zusammenhang mit T. verrucosum-Immumtät und Abheilung beim Kalb vorliegt. Resumen Por medio de biposias cutáneas secuenciales, medida de anticuerpos y exámen microscópico de presencia de hongos, se estudió la respuesta inmunitaria de tipo humoral y celular producida por la infección experimental con T. verrucosum. El exámen histopatológico reveló la presencia de agregación de linfocitos y otras células inflamatorias procedentes de la dermis de los puntos infectados. La tintura por medio de inmunoperoxidasa de las secciones congeladas, con preparaciones monoclonales de anticuerpos, demostró un aflujo de macrófagos BoCD4 + y BoCD8 + linfocitos y linfocitos, Tαδ, desde el quinto hasta el día 33 la infección. También se observó un aflugo moderado de linfocitos B. La tintura aúrica de proteina coloidal G reveló la presencia de inmunoglobulinas en al dermis y capas superficiales de la epidermis. Los anticuerpos específicos para la especie Trichophyton aparecieron entre los días 33 y 55. El exámen microscópico de los tejidos afectados demostró un incremento, de los elementos füngicos T. verrucosum (micelio y esporas exótricas) desde los días 19 al 55. Los elementos fúngicos en áreas infectadas no disminuyeron hasta después del establecimiento de ambos tipos de respuesta inmunitaria, humoral y celular. Estas respuestas implican que la combinación de ciertos fenómenos de inmunidad celular y humoral, están relacionados con la desaparición y la inmunidad de la infección producia por T. verrucosum en el ternero.     

Association of tumor budding with tumor-infiltrating lymphocytes and prognosis in breast cancer patients

AIM: To investigate the relationship of tumor budding with clinicopathologic parameters, tumor-infiltrating lymphocytes (TILs) of tumor microenvironment and the prognosis in breast cancer patients.METHODS: A total of 178 HE section samples were collected from the breast cancer patients treated with surgery in the First Affilated Hospital of Jinan University during Jan. 2012 to Dec. 2016. The tumor budding and stromal tumor-infiltrating lymphocytes were observed under light microscope. The correlation of tumor budding with the clinicopathologic status and TILs were analyzed by χ² test. Kaplan-Meier survival analysis and Log-rank test were used to estimate the disease-free survival (DFS) and overall survival (OS).RESULTS: High tumor budding level was associated with more positive lymph nodes, higher grade, and more lymphovascular invasion. In addition, the patients with higher tumor budding level showed fewer TILs, while the patients with lower tumor budding level had more TILs. Furthermore, the patients with higher tumor budding level had a worse disease-free survival and overall survival than those with lower tumor budding level.CONCLUSION: Tumor budding is significantly associated with adverse clinicopathological characteristics of breast cancer and negatively correlated with TILs. Therefore, tumor budding may serve as a potential biomarker to predict the prognosis of breast cancer.     

互叶白千层油(茶树油)对肉鸡小肠黏膜组织结构及肠道免疫细胞的影响

选用1日龄的雌性岭南黄肉鸡99只,随机分为3组,分别为对照组、试验Ⅰ组和试验Ⅱ组,每组33只。对照组常规饲养,试验Ⅰ组和Ⅱ组分别在饮水中添加0.012 mL/L和0.020 mL/L的茶树油,试验至45日龄结束。解剖取十二指肠、空肠、回肠进行HE染色和PAS染色,观察小肠黏膜形态、上皮内淋巴细胞及杯状细胞数目。结果显示,2个试验组与对照组相比,各段小肠的绒毛长度增加,肠壁变薄,隐窝深度变浅,肠绒毛高度/隐窝深度比值增加;2个试验组的上皮内杯状细胞和淋巴细胞的数目高于对照组。结果表明,茶树油能够改善肉鸡小肠黏膜结构,增加小肠上皮内免疫细胞数量,提示茶树油对肉鸡的肠道吸收和肠道免疫功能具有增强作用。     

沙冬青种子总黄酮活性成分分析及对小鼠免疫功能的影响

本研究旨在通过高效液相色谱-质谱联用（LC-MS）和高效液相色谱（HPLC）技术对沙冬青种子总黄酮主要活性成分进行分析及含量测定,并探索沙冬青种子总黄酮对小鼠免疫功能的影响。采用LC-MS分析沙冬青种子总黄酮主要活性成分,在HPLC优化条件下用标准品对主要活性成分的含量进行测定;采用MTT法和ELISA试剂盒检测沙冬青种子总黄酮对小鼠脾淋巴细胞增殖和IL-2、IL-4生成的影响,以及总黄酮对肝脏KCs细胞的增殖和NO、NOS含量的影响;运用定量溶血分光光度法和血清溶血素HC₅₀测定法检测不同浓度沙冬青种子总黄酮对小鼠抗体生成细胞和血清溶血素的影响。结果显示,沙冬青种子总黄酮中主要活性成分为芒柄花素、7,3'-二羟基-4'-甲氧基黄酮、黄豆黄素和穗花杉双黄酮,其中,芒柄花素含量高达52.48%,其次为7,3'-二羟基-4'-甲氧基黄酮,含量为11.20%。在2.5~100 μg/mL总黄酮作用下,沙冬青种子总黄酮对脾淋巴细胞的增殖和细胞因子的产生具有明显的促进作用,尤其在20 μg/mL时脾淋巴细胞增殖率和IL-2的分泌量分别高达217.62%和159.661 pg/mL,与空白对照组相比差异极显著（P<0.01）;在40 μg/mL时,脾淋巴细胞IL-4的分泌量高达149.274 pg/mL,极显著高于空白对照组（P<0.01）。沙冬青种子总黄酮对KCs细胞的增殖及NO和NOS分泌也具有明显促进作用,在60 μg/mL时,与空白对照组相比,KCs细胞增殖率达67.77%,NO和NOS的分泌量分别达180.106和29.942 μmol/L（P<0.01）。此外,沙冬青种子总黄酮能明显促进小鼠体内抗体生成细胞和血清溶血素含量的增加。综上表明,沙冬青种子总黄酮主要活性成分为芒柄花素和7,3'-二羟基-4'-甲氧基黄酮。沙冬青种子总黄酮对小鼠免疫活性细胞的增殖、相关免疫细胞因子和抗体的产生与释放等都具有显著的促进作用。     

牛流行热病毒灭活疫苗免疫后牛外周血淋巴细胞亚类分布的研究

对牛流行热病毒灭活疫苗免疫并攻每后，牛外周血淋巴细胞亚类的变化进行了研究。结果表明，灭活苗免疫后，牛的CD4^ 显著升高，可能与参与辅助B淋巴细胞合成抗体有关，攻毒后，γδT细胞均显著上升，免疫组在攻毒后3周仍保持在相当的高水平，IL－2Rα阳性细胞在攻毒后高热期也有显著升高，而CD8^ 细胞没有明显变化。     

Influence of sodium diethyldithiocarbamate (Imuthiol) on lymphocyte function and growth in weanling pigs

Mitogen-stimulated lymphocyte proliferation, delayed-type hypersensitivity (DTH) reactions, interleukin-2 (IL-2) production, and growth performance were evaluated in 3-week-old pigs treated with imuthiol. Lymphocyte proliferative responses to Con A and PWM were reduced (P < 0.05) in pigs treated with imuthiol at 25 mg/kg; PHA proliferative responses were not influenced by imuthiol treatment. Imuthiol at 2.5 mg/kg and 25 mg/kg lowered IL-2 production when compared to saline-treated controls. Delayed-type hypersensitivity responses to PHA were higher in 25 mg/kg imuthiol-treated pigs; however, 2.5 mg/kg imuthiol-treated pigs had lower DHT reactions. Imuthiol at 2.5 mg/kg and 25 mg/kg reduced (P < 0.05) average daily feed intake. These data suggest that in vivo imuthiol treatment in pigs lowers lymphocyte proliferative responses, IL-2 production, and growth performance.     

Direct and indirect lymphocyte participation in the immunity and immunopathology of tropical canine pancytopenia—A review

Erlichia canis, a rickettsial pathogen which infects monocytes, induces generalized lymphocyte activation. Activated T lymphocytes differentiate into effector cells capable of destroying infected and uninfected monocytes and platelets. Activated B lymphocytes differentiate into plasma cells associated with plasmocytosis, hypergammaglobulinemia with high levels of specific antibody and a platelet migration inhibition factor. These effector mechanisms, aimed at parasite destruction, contribute to the pathogenesis of acute tropical canine pancytopenia and do not completely eliminate the etiologic agent. A carrier state of ‘infection-immunity’ exists between the immunocompetent host and virulent parasite. E. canis has evolved mechanisms of ‘tolerant-symbiosis’ with hostile macrophages and other effector cells in the immune host.