BAT mAb induces lymphopoiesis in nude mice

The athymic nude mouse provides a powerful tool in the study of human tumors, as it enables growth of human tumors due to deficiencies in T cell functions. However, deficiencies in T cell functions might limit research on efficacy of immune modulators in cancer immunotherapy. BAT mAb mediates its anti-cancer activity through modulation of the immune system that involves both NK and T cells. We analyzed lymphocyte populations in blood 5 and 14 days following the injection of BAT antibody alone or following engraftment of human colon carcinoma cells. Our results demonstrate that BAT injection induced lymphopoiesis in the nude mouse. Percentage of CD3 cells increased up to 24%, CD4 cells up to 20% but no increase was found in CD8 T cells in BAT-injected nude mice. Injection of BAT 12 days post-tumor engraftment propagated CD3, CD4 and CD8 cells seen in the blood 5 days later but not seen in the blood 14 days post-BAT injection. It is possible that this decrease is associated with migration of the lymphocytes from the blood to the tumor sites in the livers. The percentage of CD56-positive NK cells increased (up to 18%) by BAT administration alone or post-tumor injection. The presence of tumors alone did not induce lymphopoiesis in the nude mice. Propagation and lymphopoiesis by BAT mAb might have future clinical implications.     

Restoration of hemopoiesis and immunopoiesis with small intestinal epitheliocytes in lethally irradiated mice

We studied hemopoiesis-and immunopoiesis-restoring activity of small intestinal epitheliocytes in lethally irradiated mice. The populations of peripheral blood cells and parameters of humoral and cellular immune response in lethally irradiated mice returned to normal 6 months after transplantation of cells from the small intestinal epithelial layer. Study of the distribution of intestinal cells in the body after transplantation showed longterm persistence of the donor cells in tissues of lethally irradiated animals. These data attest to high hemopoietic potential of small intestinal cells. __________ Translated from Kletochnye Tehnologii v Biologii i Medicine, No. 2, pp. 88–91, April, 2007     

Role of hyaluronidase in the regulation of hemopoiesis

We evaluated the possibility of hemopoiesis stimulation with hyaluronidase. The response of the blood system depended on the dose of this enzyme. Functional activity of hemopoietic precursor cells increased under the influence of hyaluronidase in a dose of 20 arb. units, which was accompanied by an increase in the secretion of hemopoietins by adherent myelokaryocytes and elevation of hemopoietic activity in blood plasma. The number of erythrokaryocytes and mature neutrophilic granulocytes in the bone marrow, as well as the count of reticulocytes and neutrophils in the peripheral blood increased under these conditions. Administration of hyaluronidase in a high dose (100 arb. units) led to uncoupling of proliferation and differentiation of hemopoietic precursors and produced no considerably changes in the blood. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 144, No. 12, pp. 690–695, December, 2007     

Transplantation of ACTH-secreting pituitary tumor cells in athymic nude mice

Summary Chronic excess of glucocorticoids results in cushing's syndrome in humans. A common cause of excess cortisol secretion is the presence of an adrenocorticotropin secreting pituitary tumor which stimulates the adrenal cortex to produce excess glucocorticoids. ACTH-secreting AtT-20 mouse pituitary cells transplanted subcutaneously in oestrogenized athymic nude mice form tumors rapidly. Six weeks after receiving the tumor transplants, the mice weighed 45% more than normal mice due to the increase in body fat. The tumor-bearing mice exhibit the familiar buffalo hump appearance due to the abnormal distribution of body fat. The adrenal glands of the tumor-bearing animals are enlarged due to hypertrophy of the zona fasciculata. The foamy looking fasciculata cells in normal mice were converted to dense, eosinophilic cells in the tumor-bearing mice. Transplantation of normal pituitary glands to athymic nude mice with or without oestrogen treatment did not produce these morphological changes. The experimental model described here may be useful for future studies of Cushing's syndrome.Supported by Medical Research Council of CanadaScholar, Medical Research Council of Canada     

Role of Thy-1,2+ cells in hemopoiesis regulation during hypoxia

We studied the role of Thy-1,2⁺ cells in the regulation of hemopoiesis during oxygen deficiency of different genesis. These cells of the hemopoiesis-inducing microenvironment play an important role in the compensatory and adaptive reactions of the blood system to hypoxia. Thy-1,2⁺ cells directly or indirectly (via interaction with adherent myelokaryocytes) stimulated hemopoietic precursors. The effect of these cells on committed erythroid precursors was most pronounced. __________ Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 141, No. 5, pp. 491–494, May, 2006     

Unusual cytotoxic activities of thymus-independent, self-antigen-specific CD8(+) T cells

We compared the cytotoxic activities of thymus-dependent and thymus-independent CD8(+) T cells. Thymus-dependent CD8(+) T cells, which are foreign antigen specific, acquired cytotoxic activity to tumor cells with a basal dose of the antigen peptides and to hybridoma cells expressing anti-TCR mAb only after differentiation into effector cytotoxic T lymphocytes (CTL). In contrast, thymus-independent CD8(+) T cells, which have been shown to be self-antigen specific, never showed cytotoxic activity to the target cells with a basal dose of the self-antigen peptide, while they could lyse hybridoma cells expressing anti-TCR mAb even without prior antigenic stimulation. Furthermore, the ex vivo cytotoxic activity of thymus-independent CD8(+) T cells was also observed against the target cells with high doses of the antigen peptides, which were not lysed by freshly isolated thymus-dependent CD8(+) T cells. Thus it is revealed that thymus-independent, self-antigen-specific CD8(+) T cells already acquire mature CTL functions in situ but have an increased threshold of TCR-mediated signaling for activation. These differences in cytotoxic activities between thymus-dependent and thymus-independent CD8(+) T cells suggest distinct roles of the two subsets of CD8(+) T cells in vivo.     

Functionally active CD8alphabeta+ TCRgammadelta intestinal intraepithelial lymphocytes in athymic nu/nu mice

Murine intestinal intraepithelial lymphocytes (IEL) encompass a high proportion of TCRgammadelta cells. A vast majority of these TCRgammadelta IEL express CD8alpha, but not CD8beta (CD8alphaalpha homodimer), and are considered to develop in intestinal epithelial layers independently of a functional thymus. Here we show that TCRgammadelta cells expressing both CD8alpha and CD8beta (CD8alphabeta heterodimer) appear in athymic nu/nu mice, although their appearance is random. The IEL comprising CD8alphabeta(+) TCRgammadelta cells expressed pronounced cytolytic and IFN-gamma-producing activities after TCRgammadelta ligation, which were markedly stronger than activities of IEL lacking CD8alphabeta(+) TCRgammadelta cells. Purified CD8alphabeta(+) TCRgammadelta cells expressed strong cytolytic activities and produced large quantities of IFN-gamma after TCR engagement. CD8alphabeta(+) TCRgammadelta cells were also identified among IEL from euthymic C57BL/6 mice, although their abundance varied among individual animals. However, cytolytic and IFN-gamma-producing activities in euthymic C57BL/6 mice were markedly lower than those in athymic nu/nu mice. Our findings suggest that CD8alphabeta(+) TCRgammadelta cells can develop in the intestine independently of a functional thymus/thymic epithelial cells and that they perform biological functions in situ.     

Effect of a highly purified factor from the thymus on cellular and humoral indices of immunity in thymectomized mice

In experiments on thymectomized adult CBA mice the effect of a homogeneous factor of polypeptide nature from the thymus, with mol. wt. about 5000 (thymarin-III) on the cellular and humoral indices of immunity was studied in animals. Thymectomy in animals was shown to sharply reduce the number of T-cells in the spleen. Correspondingly, the ability of the mice to produce both IgM- and IgG-antibody-forming cells and humoral antibodies against a thymus-dependent antigen (sheep's red blood cells) was sharply inhibited in the mice. Subcutaneous injection of thymarin-III in a dose of 1 g/kg into the animals daily for 7 days completely restored the T-cell population of the spleen and restored the normal immunologic reactivity of the animals.Department of Microbiology and Immunology, Institute of Experimental Medicine, Academy of Medical Sciences of the USSR, Leningrad. (Presented by Academician of the Academy of Medical Sciences of the USSR V. I. Ioffe.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 7, pp. 51–53, July, 1978.     

Achievement of cellular immunity and discordant xenogeneic tolerance in mice by porcine thymus grafts

Specific cellular immune tolerance may be essential for successful xenotransplantation in humans. Thymectomized (ATX), T and NK ceil-depleted immunocompetent mice grafted with xenogeneic fetal pig thymic and liver tissue (FP THY/LIV) result in efficient mouse thymopoiesis and peripheral repopulation of functional mouse CD4^ T cell.Very importantly, the reconstituted mouse T cells are specifically tolerant to pig donor antigens. Studies demonstrated that porcine MHCs mediated positive and negative selection of mouse thymocytes in FP THY grafts, whereas mouse MHCs were involved in negative selection in grafts. Therefore, T cell tolerance to xenogeneic donor antigens could be induced by grafting donor thymus tissue. Xenogeneic thymic replacement might have a potential role in the reconstitution of cellular immunity in patients with AIDS or other immunodeficiencies caused bv thvmus dvsfunction.     

人胰腺癌裸鼠移植瘤浸润和转移方式的研究

为了探讨胰头癌浸润及转移规律，我们用自建的三株人胰头癌裸鼠移植瘤，接咱在鼠背部、腹腔及抓垫皮下。结果表明：三株移植瘤背部接种均以局部浸润为主，可经淋巴疲乏转移至局部淋巴结（ＰＣＴ－３转移率为３３％；ＰＣＴ－４为４０％；ＰＣＴ－５为３３％），无远处淋巴结和（或）脏器转移；腹腔接种及抓垫接种与背部皮下接种结果相似。肿瘤的生长速度和浸润及转移的能力与肿瘤分化程度有一定关系。     

Analysis of T cell repertoire and function in mice transgenic for the human V{beta}3 TCR

We have constructed mice containing the human V_ß3TCR gene from the influenza virus haemagglutinin specific humanCD4⁺ T cell clone HA1.7. Similar cell yields were obtained fromtransgenic and non-transgenic lymphoid tissue, with normal levelsof T cells and with no unusual bias of the CD4 or CD8 subpopulations.Immunostaining and FACS analysis of transgenic thymocytes, spleen,and mesenteric lymph nodes revealed that the majority of T cellsexpressed the human V_ß3 TCR on the cell surface. Smallnumbers of cells expressing murine TCR_ßchain werealso detected. Polymerase chain reaction analysis revealed thatan extensive V TCR repertoire was used in the human V_ß3transgenic mice. Lymphocytes from the spleen and bmesentericlymph nodes of transgenic mice were assessed for functionalactivity in vitro. Isolated cells were stimulated with mitogenor superantigen, as well as directly through the TCR-CD3 complex,and their ability to proliferate and secrete lymphokines analysed.Cells from transgenic mice responded well after stimulationwith phytohaemagglutinin, concanavalin A, anti-CD3 antibody,anti-CD3 antibody with phorbol ester, and Staphylococcus aureusenterotoxin B, and also showed alloreactivity in a mixed lymphocytereaction. Minimal levels of response were detected after stimulationwith murine TCRß antibody. Together, these data suggestthat a human TCRß chain is able to associate witha murine TCR chain, to form a fully functional surface TCR-CD3complex.     

Adoptive transfer of nontransgenic mesenteric lymph node cells induces colitis in athymic HLA-B27 transgenic nude rats

HLA-B27 transgenic (TG) rats develop spontaneous colitis when colonized with intestinal bacteria, whereas athymic nude (rnu/rnu) HLA-B27 TG rats remain disease free. The present study was designed to determine whether or not HLA-B27 expression on T cells is required for development of colitis after transfer of mesenteric lymph node (MLN) cells into rnu/rnu HLA-B27 recipients. Athymic nontransgenic (non-TG) and HLA-B27 TG recipients received MLN cells from either TG or non-TG rnu/+ heterozygous donor rats that contain T cells. HLA-B27 TG rnu/rnu recipients receiving either non-TG or TG MLN cells developed severe colitis and had higher caecal MPO and IL-1beta levels, and their MLN cells produced more IFN-gamma and less IL-10 after in vitro stimulation with caecal bacterial lysate compared to rnu/rnu non-TG recipients that remained disease free after receiving either TG or non-TG cells. Interestingly, proliferating donor TG T cells were detectable one week after adoptive transfer into rnu/rnu TG recipients but not after transfer into non-TG recipients. T cells from either non-TG or TG donors induce colitis in rnu/rnu TG but not in non-TG rats, suggesting that activation of effector T cells by other cell types that express HLA-B27 is pivotal for the pathogenesis of colitis in this model.     

Role of Thy-1,2<Superscript>+</Superscript> cells in the regulation of hemopoiesis during severe hypoxia

We studied the state of the bone marrow Thy-1,2⁺ cell pool under conditions of severe hypoxia. T cell mechanisms of hemopoiesis regulation are preserved under conditions of severe oxygen deficiency due to changes in functional properties of Thy-1,2⁺ cells. We revealed an indirect (mediated through cooperation with adherent myelokaryocytes) stimulating effect of Thy-1,2⁺ cells on erythroid precursors as well as direct and indirect feeder effects of these cells on granulocyte-monocyte precursors. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 143, No. 1, pp. 23–27, January, 2007     

Hemopoiesis in the Thymus

The presence in the thymus of hemopoietic cells other than thymocytes has been known for many years, but the extent of the hemopoietic activity of the thymus and the possible functional implications have only recently begun to receive much attention. This review summarizes the literature in this field, especially in the light of current cytokine and thymic-factor knowledge, and includes clinical relevance where possible.     

Spontaneous development of autoimmune uveoretinitis in nude mice following reconstitution with embryonic rat thymus.

This paper describes the spontaneous occurrence of an autoimmune uveoretinitis in nude (nu/nu) mice reconstituted when 4 weeks old by the grafting of rat embryonic thymus. The uveoretinitis was characterized histologically by progressive loss of the photoreceptor layer, observed in 4.0, 17.6, 42.9% and 71.4% of such mice at 3, 5, 7 and 12 months of age, respectively. Mice with uveoretinitis were shown to have serum IgG antibody reactive by indirect immunofluorescence with retinal photoreceptors, and with interphotoreceptor retinoid-binding protein (IRBP), but not retinal S-antigen, by immunoblotting and ELISA. A uveoretinitis could be adoptively transferred to syngeneic ungrafted nude mice by splenic CD4+ T cells from diseased animals. This is the first experimental model of a (T cell mediated) autoimmune uveoretinitis which develops spontaneously and which is not dependent upon deliberate sensitization with retinal antigens and adjuvants.     

细胞外基质受体α-Dystroglycan影响胸腺细胞分化发育的可能机制

目的研究细胞外基质受体alpha-Dystroglycan(α-DG)对胸腺细胞分化的影响及机制。方法摘取15日胚龄鼠胸腺小叶进行体外器官培养。将α-DG抗体、对照抗体或培养液滴加在胸腺小叶上。FACS(Fluorescence-activated cell sorting)分析胸腺细胞表面分子CD4、CD8、CD95和CD69等的表达。结果α-DG中和抗体能明显抑制胸腺细胞分化，显示胸腺双阴性细胞比例从对照组的26．5％增高到实验组的71．6％，双阳性细胞和CD8单阳性细胞比例则显著下降，分别从39．8％和20．7％下降到7．5％和6．8％，CD4单阳性细胞比例则无明显变化；同时胸腺细胞数目明显减少；CD95、CD69的表达水平随α-DG中和抗体的持续存在呈现显著升高。结论α-DG通过参与胸腺细胞的活化和凋亡活动影响胸腺细胞的发育。     

{gamma}{delta} T cells promote CD4 and CD8 expression by SCID thymocytes

Molecular studies of the TCR, which is expressed by a minorsubpopulatlon of T lymphocytes in all vertebrate species, havedefined a subset which expresses a receptor with extreme junctionaldiversity and a second subset, most commonly found in eplthella,which expresses a receptor of very limited diversity. In thedeveloping murine thymus, T cells appear in an ordered sequenceof specific v rearrangements, V3V, 1 on day 14, V2V1 on day17, and subsequently V4V5, V6, or V7. We demonstrate that thetransfer of expanded populations of cells from newborn thymusand cell lines expressing the invariant V3V1 receptor into SCIDmice, which lack T and B cells, results in the appearance ofCD3^–CD4⁺CD8⁺ thymocytes. Thus, one role of the early appearingV3V1 T cells in thymlc development in vivo is to promote CD4and CO8 surface expression on precursor cells.     

李斯特菌感染对小鼠APC活性的影响

目的：通过对转基因鼠脾T细胞的分化研究,探讨感染初期APC的活性。方法：细胞因子ELISA测定转基因鼠脾T细胞产生的IFN-γ及IL-4,FACS鉴定T细胞为TCR-TG的T细胞（Vβ8.2品系）。结果：感染鼠的APC诱导TG鼠的T细胞向Th1方向分化,未感染鼠的APC诱导TG鼠的T细胞向Th2方向分化,IL-12和IL-4可影响APC对T细胞分化的诱导。结论：感染和外来细胞因子影响APC的提呈诱导活性。