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.     

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。     

Cytochemical properties of chicken blood cells resembling both thrombocytes and lymphocytes

Generally accepted criteria were used to identify typical nucleated thrombocytes and typical small lymphocytes in chicken-blood smears subjected to modified-Wright staining. Other cells, here referred to as "intermediate cells," were difficult to classify because in some aspects they resembled thrombocytes while they also had features typical of small lymphocytes. The "intermediate cells" had small, round or oval nuclei with coarsely condensed chromatin, characteristic of both thrombocytes and small lymphocytes. In addition, "intermediate cells" had moderately abundant cytoplasmic volumes, typical of thrombocytes but blue cytoplasm lacking both granules and vacuoles, which is characteristic of small lymphocytes. It made little difference to the thrombocyte count whether these cells were classified as thrombocytes or small lymphocytes; however, this decision made a substantial difference to the lymphocyte count in some chicken-blood smears. Most "intermediate cells" (351 of 410 cells examined) were nonfluorescent after treatment with formaldehyde gas. Furthermore, most "intermediate cells" failed to acquire characteristic pigments when subjected to either Grimelius staining (179 of 204 cells examined) or periodic acid-Schiff staining (173 of 206 cells examined). Typical small lymphocytes reacted in the same way, failing to fluoresce after gaseous formaldehyde treatment (65 of 65 cells examined) and failing to react during Grimelius staining (41 of 44 cells examined) or periodic acid-Schiff staining (21 of 21 cells examined). In contrast, almost all typical thrombocytes became fluorescent in response to gaseous formaldehyde (709 of 718 cells examined) and gave positive reactions when subjected to Grimelius staining (381 of 382 cells examined) or periodic acid-Schiff staining (322 of 326 cells examined). These findings suggested that "intermediate cells" should be classified as lymphocytes in differential cell counts.     

Identification of feline blood B and T lymphocytes by cell electrophoresis.

The electrokinetic properties of feline blood lymphocytes isolated by centrifugation over Ficoll-Isopaque gradient were investigated. A biphasic electrophoretic mobility (EPM) distribution was regularly observed with a low-mobility (LM) population (mean EPM: 0.82) accounting for 32% of blood lymphocytes and a high-mobility (HM) population (mean EPM: 1.09) representing 68% of blood lymphocytes. Following fractionation on nylon-wool columns, lymphocytes with B-cell properties (64% sIg⁺; 9% guinea pig erythrocytes (GPE)-rosette⁺, PHA and Con A unresponsive) were enriched in the adherent fraction and belonged mainly (78%) to the LM population. In contrast, lymphocytes with T properties (5% sIg⁺, 42% GPE-rosette⁺, PHA and Con A responsive) were recovered in the effluent fraction and comprised 84% of HM elements.Thus, in cat blood, LM lymphocytes are likely to represent in majority B cells and HM lymphocytes T cells. This indicates that cell electrophoresis provides an interesting mean for differentiating B and T cells in the cat.     

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)。     

Tyrosine Ameliorates Some of the Clinical,Biochemical and Haematological Effects of Acute Stress Associated with Transportation of Desert Sheep

We studied some clinical, biochemical and haematological variables in Desert (Najdi) sheep acutely stressed in the course of individual road transportation, and the influence thereon of pretreatment with tyrosine. Transportation for 30 min resulted in variable but statistically insignificant increases in heart, pulse and respiratory rates. It also caused significant increases in the plasma concentration of cortisol (from 43.5 to 101.7 mmol/L) and glucose (from 3.1 to 4.5 mmol/L), and a decrease in that of magnesium (from 0.85 to 0.72 mmol/L). The endogenous thiocyanate level was unaffected. The transportation stress also decreased the haematocrit (PCV) and the number of lymphocytes, and increased the concentration of haemoglobin. Pretreatment of sheep with tyrosine at a dose of 100 mg/kg by the intravenous route significantly ameliorated the stress-induced clinical, biochemical and haematological changes. The treatment caused no overt adverse effects.     

New gene-based approaches for an AIDS vaccine

Vaccine approaches against AIDS have focused on inducing cellular immune responses, since many studies revealed the role of T cell responses in the control of human immunodeficiency virus or simian immunodeficiency virus (SIV) infections. The experimental infection of rhesus macaques with SIV or chimeric SHIV is routinely used as a model for AIDS. In such models, DNA immunization is a tool to elicit specific T cell responses and to study their protective efficacy. DNA immunogenicity in primates depends on parameters such as level of antigen expression, choice of the antigen among SIV proteins, use of fusion proteins, route of immunization, and addition of adjuvants. Recent results suggest that priming with DNA and boosting with attenuated recombinant viral vectors, each expressing corresponding SIV antigens, leads to improved specific immunity and, in some cases, affords protection against pathogenic challenge. After preclinical evaluations, DNA has entered clinical trials for a therapeutic or prophylactic gene-based AIDS vaccine.     

Seasonal atopic dermatitis in dogs sensitive to a major allergen of Japanese cedar (Cryptomeria japonica) pollen

Three dogs were examined because of episodes of recurrent pruritic dermatitis in the spring, the season of Japanese cedar (Cryptomeria japonica, CJ) pollination in Japan. The dogs were shown to be sensitive to CJ pollen allergen using intradermal testing and antigen-specific IgE measurement. Fluorometric enzyme-linked immunosorbant assay (ELISA) showed increased concentrations of IgE specific to Cry j 1 and a negative result for Cry j 2 in the three dogs. The concentrations of IgE specific to Cry j 1 during the season of CJ pollination were higher than the concentrations found during the off-season in all the dogs, and the variation in the concentrations correlated with the variation in clinical signs. Peripheral blood mononuclear cells showed apparent proliferative responses to crude CJ pollen antigen and Cry j 1 during CJ pollination season. These findings indicated that Cry j 1 was the major allergen recognized by IgE and lymphocytes and resulted in the development of type I hypersensitivity to CJ pollen allergen in these atopic dogs.