Protection of Lethally Irradiated Mice by Spleen Cells from Neonatally Thymectomized Mice

Long-lived, immunologically vigorous (C3H(f) x A(f))F(1) hybrids were produced after lethal irradiation by administration of spleen cells from C3H(f) or syngeneic donors. Further, neonatally thymectomized C3H(f) or A(f) strain donors reconstituted irradiated C3H(f) or (C3H(f) x A(f))F(1) hosts. In addition, C3H(f) spleen cells from nonthymectomized 10- to 15-day-old donors protected irradiated hybrid mice, but A(f) cells of young mice as well as of older mice produced graft-versus-host reaction and early death in irradiated C3H(f) or (C3H(f) x A(f))F(1) hybrids.Abrogation of secondary disease by treatment of irradiated mice with spleen cells from allogeneic neonatally thymectomized mice is possibly attributable to diminished immunologic competence of the cells grafted, followed by the development of immunological tolerance of the donor cells. Donor cells, receiving thymus influence in the recipient host after transplantation, could explain the long-lived immunologically vigorous radiation chimeras that did not experience graft-versus-host reactions. The findings of this study help to understand the differential susceptibility of A(f) and C3H(f) mice to development of tolerance to one another's antigens observed in prior investigations.It appears that, in these mice, the host thymus influences the maturation of the spleen cells from young mice or from neonatally thymectomized mice. However, this influence was often greater in mice given 767 rads than in those given 1046 rads. This differential influence is possibly attributable to irradiation damage to the thymus produced by the higher dose of irradiation. Spleen cells from neonatally thymectomized mice can be differentiated and expanded by the thymus of the host. The differential susceptibility of T(1), early differentiation stages, of thymus-dependent lymphocytes and T(2), late differentiation stages, of thymus-dependent lymphocytes to tolerance induction and immunostimulation, respectively, are proposed as the bases for these otherwise paradoxical influences.     

Antihypertensive effect of thymectomy in Lyon hypertensive rats Vascular reactivity,renal histology,and sodium excretion

The aim of this study was to search for the possible mechanisms involved in the antihypertensive effect of neonatal thymectomy that we previously observed in Lyon hypertensive (LH) rats. To that end, we studied in LH and normotensive control (LN) rats the consequences of neonatal thymectomy on vascular reactivity, renal structure, and pressure-natriuresis. The increase in pressor responses to angiotensin I and phenylephrine noted in LH rats as compared to LN animals was abolished by neonatal thymectomy. Histological study showed that kidneys from LH rats exhibited arterial wall hypertrophy, segmental hyalinization of the glomeruli, and were infiltrated by mononuclear cells. All these features of kidney injury were reduced in neonatally thymectomized LH rats. Lastly, the responses of isolated perfused kidneys from LH rats to stepwise reductions in renal perfusion pressure differed from those of LN rats by decreased renal perfusion flow and natriuresis. Neonatal thymectomy tended to improve sodium excretion in parallel with a slight decrease in renal vascular resistances. It is concluded that the normalization of vascular responsiveness to vasoconstrictor factors, the alleviation of renal lesions and, to a lesser extent, the moderate improvement of pressure natriuresis may account, at least in part, for the antihypertensive effect of neonatal thymectomy in LH rats.     

The parietal cell autoantigens recognized in neonatal thymectomy-induced murine gastritis are the alpha and beta subunits of the gastric proton pump [corrected]

Murine autoimmune gastritis, induced by neonatal thymectomy, bears a striking similarity in pathology to the human autoimmune disease, pernicious anemia. Autoantibodies to parietal cells are found in both murine and human diseases. Monoclonal immunoglobulin G autoantibodies, obtained from neonatally thymectomized mice, have previously been shown to recognize two groups of gastric parietal cell antigens. In the present study, it is shown that two of these monoclonal autoantibodies, designated 1H9 and 2B6, are directed against the alpha subunit and beta subunit, respectively, of the gastric hydrogen-potassium-stimulated adenosine triphosphatase (H+,K(+)-ATPase; proton pump). Monoclonal antibody 1H9 showed reactivity by immunoblotting with a 95-kilodalton component of dog gastric tubulovesicular membranes and with a fusion protein containing the hydrophilic domain of the alpha subunit of the H+,K(+)-ATPase. Monoclonal antibody 2B6 reacted by immunoblotting with the 60-90-kilodalton glycoprotein (beta subunit) of the tomato lectin-purified dog H+,K(+)-ATPase and with the 60-90-kilodalton autoantigen purified with human parietal cell autoantibodies. Monoclonal antibody 2B6 also reacted with the deglycosylated 35-kilodalton core protein of the tomato lectin-purified 60-90-kilodalton beta subunit and of the purified 60-90-kilodalton autoantigen. Parietal cell autoantibody-positive sera from 20 mice with experimentally induced gastritis showed reactivity predominantly with the alpha and/or beta subunit of the gastric H+,K(+)-ATPase. Therefore, it is concluded that the major molecules targeted by parietal cell autoantibodies from mice with neonatal thymectomy-induced murine autoimmune gastritis and from humans with pernicious anemia are identical.     

The role of NZB/NZW F1 thymus in experimental tolerance and auto-immunity

The role of the NZB/NZW F₁ (B/W) thymus was studied with respect to tolerance to ultracentrifuged bovine γ globulin (BGG) and poly I· poly C, and spontaneous anti-DNA antibody formation. Thymectomy alone had little effect on the induction or escape from tolerance. An adult B/W thymus could not prevent toleranceinduction in young B/W mice. However, a young B/W thymus could facilitate the induction of partial tolerance in older B/W mice under appropriate conditions. These results suggest that the adult B/W thymus may be functionally deficient relative to the young thymus. Neonatal thymectomy accelerated the autoimmune disease, suggesting that the young B/W thymus has some normal regulatory function. Thymectomy at 6 weeks of age did not accelerate autoimmunity. A young B/W thymus, but not an older B/W thymus reversed the accelerated disease of neonatally thymectomized B/W mice. This again suggests that the older B/W thymus is functionally deficient.     

枯否细胞对同种异系抗原的捕获与门静脉免疫耐受的关系

目的探讨肝脏及肝脏内Kupffer细胞在同种异系抗原门静脉接种诱导的免疫耐受机制中的作用。方法以NIH／q小鼠淋巴细胞对BALB／C小鼠门静脉或腔静脉接种,一周后再行皮下接种二次,间隔时间为一周,第二次皮下接种后一周,测定供受体混合淋巴细胞增生反应(MLR)和受体对供体抗原的迟发性超敏反应(DTH)。经神经氨酸酶(neuraminidase)处理的供体淋巴细胞经受体小鼠下腔静脉接种,以及在门静脉接种前一天,预先用Kupffer细胞抑制剂三氯化钆(GdCl     

Sj26和Sj23基因转染树突状细胞联合抗血吸虫感染的研究

目的探讨Sj26和Sj23基因转染树突状细胞(DC)联合抗日本血吸虫感染保护性免疫机制。方法BALB/c小鼠耳廓分别注射Sj26和Sj23基因转染DC(A组)、Sj26基因转染DC(B组)、Sj23基因转染DC(C组)、pcDNA3转染DC(D组)、未处理DC(E组)和RPMI-1640(F组),免疫3次,间隔2周,末次免疫后2周,每鼠经皮肤感染40条日本血吸虫尾蚴。ELISA法检测血清特异性IgG抗体、干扰素-γ(IFN-γ)和白细胞介素-4(IL-4)水平,双夹心ELISA法检测脾淋巴细胞经ConA和可溶性虫卵抗原(SEA)刺激后培养上清中IFN-γ和IL-4水平,噻唑蓝(MTT)法检测脾淋巴细胞增殖情况。结果末次免疫后第2周,A组小鼠血清IgG抗体水平显著升高(P<0.001)。血清IL-4的水平,各组免疫前、后无明显变化。血清IFN-γ水平,A组小鼠免疫后明显升高(P<0.01)。经ConA和SEA刺激后,A组小鼠脾淋巴细胞诱生的IFN-γ水平显著增高,而IL-4水平显著降低(与F组比较,P<0.001)。A组小鼠脾淋巴细胞经ConA和SEA刺激后的刺激指数均高于其他各组(与F组比较,P<0.001)。结论体液免疫和细胞免疫共同参与了Sj26和Sj23基因转染DC诱导的保护性免疫作用,其中Th1型免疫应答在抗日本血吸虫感染的保护性免疫中起主要作用。     

T cells are required for coxsackievirus B1 induced murine polymyositis

Murine polymyositis (PM) induced by coxsackievirus B1 (CVB1) has been used as a model of human PM. Our study was undertaken to investigate the role of T cells in CVB1 induced PM by examining disease development in neonatally thymectomized mice. Clinical weakness and histological inflammatory myositis occurred in 42.7% of sham operated animals but only 7.7% of thymectomized mice. These experiments demonstrate the importance of T cells in the pathogenesis of this virus induced autoimmune disorder.     

慢性重型肝炎伴胃粘膜病变及门脉高压性胃病的研究

目的　观察慢性重型肝炎患者上消化道粘膜病变特点。方法　将列入观察对象的患者分为 2个组 :A组为肝硬化伴门静脉高压转重而符合慢性重型肝炎诊断者 ;B组为慢性肝炎病情转重者。同期住院的急性黄疸型肝炎病人作为对照观察。结果　A组慢重肝患者有 68 6% ( 13 1/ 191)胃体粘膜出现门脉高压性胃病 (PHG)改变 ;B组PHG为 2 4 6% ( 43 / 175 ) (P <0 0 5 ) ;A组胃粘膜病变 (GML)的检出率为 42 4% ( 81/ 191) ,与B组的45 1% ( 79/ 175 )无明显差异 (P >0 0 5 ) ;但与对照组相比差异显著 (P <0 0 1)。结论　重度PHG的发生与肝脏功能密切相关。门脉压变化影响胃粘膜微循环 ,对PHG形成有一定作用。慢性重型肝炎发生GML构成多脏器功能损伤表现之一。其发生机制可能与凝血因子合成障碍 ;门脉高压的作用 ;高胃泌素血症 ;肿瘤坏死因子(TNF)的作用有关。多种因素中某一因素可能在某一时期起主导作用。临床应重视整体治疗 ,如积极的支持疗法 ,改善肝功 ,补充凝血因子 ,纠正低氧血症 ,调节酸硷平衡。出现消化道症状及时进行胃镜检查 ,针对性处理     

Portalsystemic hemodynamic changes in chronic severe hepatitis B： An ultrasonographic study

AIM： To evaluate portalsystemic hemodynamic changes in chronic severe hepatitis B.
METHODS： Hemodynamic parameters included portal vein diameter （PVD）, portal vein peak velocity （PVPV）, portal vein volume （PW）, spleen length （SPL）, spleen vein diameter （SPVD）, spleen vein volume （SPW） and umbilical vein recanalization. They were measured by Color Doppler ultrasonography in 36 patients with chronic severe hepatitis B, compared with 51 normal controls, 61 patients with chronic hepatitis B, 46 patients with compensable cirrhosis, and 36 patients with decompensable cirrhosis.
RESULTS： In the group of chronic severe hepatitis B, PVD （12.38 ± 1.23 mm） was significantly different from the normal control, compensable cirrhosis and decompensable cirrhosis groups （P = 0.000-0.026）, but not significantly different from the chronic hepatitis group. PVPV （16.15 ± 3.82 cm/s） dropped more significantly in the chronic severe hepatitis B group than the normal control, chronic hepatitis B and compensable cirrhosis groups （P = 0.000-0.011）. PW （667.53 ± 192.83 mL/min） dropped significantly as compared with the four comparison groups （P = 0.000-0.004）. SPL （120.42 ± 18.36 mm） and SPVD （7.52 ± 1.52 mm） were longer in the normal control and chronic hepatitis B groups （P = 0.000-0.009）, yet they were significantly shorter than those in the decompensable cirrhosis group （P = 0.000）. SPW （242.51 ± 137.70 mL/min） was also lower than the decompensable cirrhosis group （P = 0.000）. The umbilical vein recanalization rate （75%） was higher than the chronic hepatitis B and compensable cirrhosis groups. In the course of progression from chronic hepatitis to decompensable cirrhosis, PVD, SPL and SPVD gradually increased and showed significant differences between every two groups （P = 0.000-0.002）.
CONCLUSION： Patients with chronic severe hepatitis B have a tendency to develop acute portal hypertension, resulting in significantly reduced portal vein     

The influence of neonatal thymectomy on the blood serum level of total lipids, NEFA, triglycerides, total cholesterol and beta-lipoproteins in rats

The time course of changes in the blood serum content of total lipids (TL), nonesterified fatty acids (NEFA), triglycerides (TG), total cholesterol (TC) and beta-lipoproteins (beta-LP), caused by neonatal thymectomy (NTE), was investigated in neonatally thymectomized and in sham-operated rats aged 15, 30, 45 days, 2, 3 and 5 months. TL were significantly increased on days 15, 30, 45 and 60 after operation; their level in 3- and 5-month-old rats was already normal. In control rats, NEFA level increased with age; their level in NTE rats was significantly higher on days 15, 30, 45 and 60 after thymectomy. Similar changes were registered in TG level. The effect of NTE was most pronounced in TC whose level was significantly higher in thymectomized rats than in control rats till the 150th day after operation. Serum concentration of beta-LP in NTE rats exceeded several times that of sham-operated rats in all age groups. The role of the thymus in the control of lipid metabolism in rats is discussed.     

Systemic bacillus Calmette-Guérin (BCG) activates natural suppressor cells.

Addition of normal C57BL/6 mouse bone marrow cells to an in vitro culture of normal C57BL/6 spleen cells and allogeneic P815-Y tumor cells inhibited the development of cell-mediated immunity. Bacillus Calmette-Guérin (BCG) enhanced the suppressive activity of these bone marrow cells as early as 2 days after its intravenous administration to donor mice and elicited similar activity in the spleen by 7 days. Concomitant with the appearance of suppressor cells in the spleen there was a decrease in bone cell number and an increase in spleen cell number. While normal spleen cells failed to inhibit immunization, spleen cells from thymectomized, irradiated, bone marrow-reconstituted mice were inhibitory. Administration of BCG further increased the suppressive activity of spleen cells in these T cell-deprived mice. From this evidence it appears that systemic administration of BCG activates natural suppressor cells in the bone marrow and elicits suppressor cells in the spleen through the migration and colonization of the spleen by bone marrow elements.     

Fas/CD95 is required for gastric mucosal damage in autoimmune gastritis

BACKGROUND & AIMS: Experimental autoimmune gastritis (EAG), characterized by a gastric mononuclear cell infiltrate, mucosal cell damage, and autoantibodies to parietal cell-associated H(+)/K(+) adenosine triphosphatase, is a model for human autoimmune gastritis that leads to pernicious anemia. Previous in vitro studies have implicated Fas/CD95 in initiating damage to gastric mucosal cells in humans and an animal model of autoimmune gastritis. Here we used 2 in vivo animal models to examine the role of Fas in the development of mucosal cell damage in autoimmune gastritis. METHODS: We initiated EAG in BALB/cCrSlc mice by neonatal thymectomy and examined for Fas expression in the gastric mucosa by immunohistochemistry. To address the in vivo relevance of Fas in mucosal injury, we examined the stomachs and sera of BALB/cCrSlc lpr/lpr mice subjected to neonatal thymectomy and BALB/cCrSlc nu/nu lpr/lpr mice transferred with lymphocytes from gastritic BALB/cCrSlc mice. RESULTS: Fas expression was up-reguiated in parietal cells of mice with EAG. Neonatally thymectomized lpr/lpr mice were resistant to developing destructive gastritis compared with heterozygous and wild-type littermates. Nu/nu Fas-sufficient mice transferred with lymphocytes from thymectomized lpr/lpr mice developed destructive gastritis. Nu/nu lpr/lpr mice transferred with lymphocytes from gastritic mice developed a nondestructive gastritis. CONCLUSIONS: The observations that Fas is up-regulated in gastric parietal cells of mice with EAG and that Fas-deficient mice are resistant to development of destructive gastritis provide compelling evidence that Fas is required in vivo for development of gastric mucosal cell damage in autoimmune gastritis.     

Involvement of suppressor cells induced with membrane fractions of trypanosomes in immunosuppression of trypanosomiasis

We showed that infection with Trypanosoma congolense in mice led to suppression of listeria-induced delayed type hypersensitivity (DTH). Mice were pre-treated with irradiated T. evansi, which caused rapid and effective suppression of DTH. A membrane fraction obtained by homogenizing T. evansi variant in a hypotonic buffer solution and centrifuging it at 150,000g produced suppression of listeria-induced DTH when injected i.p. into mice as early as 1 day before listeria immunization. Furthermore, we demonstrated by an adoptive transfer system that the suppressor cells involved in this suppression had developed in the spleen and that the activity of the splenic suppressor cells was due to the presence of a macrophage population.     

Effect of thymectomy on tumor development and on T and B lymphocytes in tumor-bearing rats.

Tumor growth and changes in T and B lymphocyte ratio in spleen, draining lymph node and peripheral blood of thymectomized, irradiated rats, reconstituted with syngeneic bone marrow transplanted at various time intervals with MC-1 fibrosarcoma cells were followed. Control nonthymectomized or "sham" operated rats were transplanted an equal dose of tumor cells. Thymectomy and irradiation reduced the numbers of T lymphocytes in all lymphoid organs, while the enhanced numbers of B cells are probably related to reconstitution with cells of syngeneic bone marrow. The time interval between thymectomy, irradiation and transplantation of tumor cells proved to be a limiting factor for tumor growth and changes in T and B cell ratio. Early transplantation of tumor cells (7 days after irradiation) resulted in an enhanced resistance to tumor development, a reduced tumor growth rate and a progressing decline in the number of T cells. If the interval between thymectomy and tumor cell transplantation lasted 4 weeks, the T cell population became partially regenerated, and tumors grew progressively in correlation with a continuing T lymphocyte depletion. The results are discussed in terms of the role of various T cell subpopulations and the significance of residual, thymectomy- and irradiation-resistant T lymphocyte population, vital for a preservation of T cell immunological functions.     

Dual colour fluorescein analysis of peripheral blood T cells in auto-immune chronic active hepatitis

Both CD4 and CD8 T cells are subdivided into two phenotypically distinct sublineages via another two T cell markers, Leu-8 and CD11b antigens. The proportions of these four T cell subsets, CD4+Leu8+, CD4+Leu8-, CD8+11b+ and CD8+11b-, were studied in patients with auto-immune chronic active hepatitis (CAH) and compared with disease controls (hepatitis B surface antigen positive chronic active hepatitis) and healthy controls. We found that the proportion of CD4+Leu8+ cells was significantly reduced compared with controls (P less than 0.01), whereas those of the other cells were almost identical in all 3 groups. The absolute number of these CD4+Leu8+ cells was also lower than that of controls (P less than 0.01). Thus, the present study suggests that a reduced number of CD4+Leu8+ cells is associated with the aberrant immune response in auto-immune CAH.     

Autoimmune hepatitis induction can occur in the liver

The priming of T cells in the liver is widely accepted. Nonetheless, it is controversial whether immune activation in autoimmune hepatitis (AIH) occurs in the liver or in the spleen. To address this issue, we splenectomized mice and induced experimental murine AIH (emAIH) with an adenovirus (Ad)‐expressing formiminotransferase cyclodeaminase (FTCD). Post‐splenectomy, the experimental mice developed emAIH to a higher extent than the control mice. In addition, splenectomized mice harboured more intrahepatic B cells and a disproportionately small number of regulatory T cells (Tregs) within a reduced T cell population at the site of inflammation. These results imply that the spleen is not the site of AIH induction. In contrast, the spleen seems to have a protective function since the pathological score was more severe in splenectomized animals. These findings have important implications for the aetiology of AIH.     

FURTHER STUDIES ON THE ROLE OF CIRCULATING LYMPHOCYTES IN THE INITIATION OF PRIMARY ANTIBODY RESPONSES TO DIFFERENT ANTIGENS

Thoracic duct cells obtained from normal (unimmunized) donors restored the primary hemolysin response of lethally irradiated or neonatally thymectomized rats to sheep red blood cells. Synergy between thoracic duct cells and bone-marrow cells was demonstrated in the irradiated hosts. However, thoracic duct cells did not restore the primary antibody response of irradiated rats challenged with diphtheria toxoid, but did restore the response of neonatally thymectomized rats. The addition of peritoneal exudate cells or bone-marrow cells to inocula of thoracic duct cells also failed to restore the response of irradiated hosts to diphtheria toxoid, although normal spleen cells restored the response. These findings indicate that the cellular events involved in the initiation of the primary antibody response to sheep red blood cells differ from those involved in the response to diphtheria toxoid.     

Leukocyte Counts of Germ-Free Neonatally Thymectomized CFW Mice

Examination of hematologic data from germ-free, germ-free thymectomized,conventional, and conventional thymectomized CFW mice revealed few statistically significant differences in hematocrit values or total leukocyte counts.Germ-free thymectomized and conventional thymectomized mice had significantly higher counts than their controls on 8 and 9 days out of the first 2 weeksfollowing thymectomy, but not after 2 weeks. A slight decrease in agranulocytes was noted in both groups of thymectomized mice only during the first2 weeks postthymectomy. With the onset of wasting disease both granulocyteand agranulocyte counts increased in the conventional thymectomized mice.

Submitted on July 12, 1966 Accepted on March 7, 1967     

On the mode of action of heterologous antileukemic sera. Growth of syngeneically transplanted leukemia cells on serum treated neonatally thymectomized mice (author's transl)]

A heterologous antiserum against a Nitrosomethylurea--induced mouse leukemia prevented the outgrowth of syngeneically transplanted leukemia cells in neonatally thymectomized mice. After subcutaneous challenge with 50 000 leukemia-ascites cells thymectomized CBA mice at the age of 8 weeks were given 5 intraperitoneal injections each of 0,1 ml of the heterologous serum. While the antileukemic serum protected 9 out of 11 mice all of the 11 mice treated with normal rabbit serum developed a tumor. The absence of the thymus was confirmed by macroscopic control and by the absence of antibodies against the injected rabbit serum. With regard to previous findings showing a cooperation of heterologous antibodies with host cells as responsible for the antileukemic effect this result indicates that the effector cells are thymus-independent.