State of cellular immunity and phagocytes in guinea pigs infected with Mycoplasma pneumoniae

The work presents the results of the study of experimental Mycoplasma pneumoniae infection in guinea-pigs. A considerable increase in the engulfment of mycoplasmas and blood leukocytes was found to occur on days 14-28 after the infection. The correlation between the degree of the engulfment of mycoplasmas by macrophages and the activity of lymphocytes in the reaction of blast-cell transformation with phytohemagglutinin and Mycoplasma antigen was observed. The peculiarities of the course of infection in sensitized animals were revealed. Virulent Mycoplasma pneumoniae strains were found to have a toxic effect on the lymphocytes and macrophages of guinea-pigs. This effect was neutralized by specific antiserum. The importance of these facts for the pathogenesis of Mycoplasma infection is discussed.     

Staphylococcal infection in guinea pigs sensitized with a commercial staphylococcal allergen

Guinea pigs, previously injected with commercial staphylococcal allergen to induce delayed hypersensitivity, were infected by the intramuscular injection of S. aureus in a nonlethal dose. For control, the animals receiving only S. aureus were used. The dynamic study of the degree of septicemia and some lymphocytic characteristics in the animals was made. The study revealed that delayed hypersensitivity did not aggravate the course of the main disease; on the contrary, it rendered protection against the subsequent infection. Increased resistance to infection was manifested by a decrease in the degree of septicemia, determined from the decreased number of colony-forming units of S. aureus in the splenic tissue as assessed by inoculation into agar, as well as from a higher level of the activation of lymphocytes as assessed by rosette formation.     

Staphylococcal infection in guinea pigs with enhanced delayed hypersensitivity induced by Staphylococcus aureus surface antigens

A comparative study was made of the relationship between the DHS phenomenon and certain staphylococcal antigens and the intensity of infectious process in sublethal infection of guinea-pigs by Staphylococcus aureus. The protective effect, which was manifested by reduced amounts of staphylococcal colonies in spleen tissues and elevated level of lymphocytes, and neutrophil activation, correlated with DHS induced by surface antigens under study (cell wall, peptidoglycan, protein A). Incomplete combination or certain antigens induced DHS but did not increase the resistance to the following infection.     

Immunological reaction of guinea-pigs following intranasal Mycoplasma pneumoniae infection and immunization with the 168 kDa adherence protein

Humoral responses to Mycoplasma pneumoniae proteins, especially the 168 kDa protein, were demonstrated by Western blotting in sera and bronchial washings of all groups of infected or immunized guinea-pigs. However, infection was not prevented by these local and systemic antibodies. Hilar lymphocytes of infected and immunized guinea-pigs were stimulated in vitro by sonicated M. pneumoniae antigen and by the 168 kDa protein. Stimulation was significantly lower in animals which had been infected twice or had been preimmunized and challenged by infection. Histologically the most severe lesions were seen in the twice-infected group followed by the preimmunized group which was subsequently infected.     

Morphological changes in experimental infection in guinea pigs under the influence of prodigiozan and S-methylmethionine]

The effect of prodigiozan and S-methylmethionine on the level of histomorphological changes in the organs and tissues of guinea pigs with experimental typhoid fever, dysentery and staphylococcal infections, as well as the effect of prednizolone with respect to dysentery and staphylococcal infections were studied. In the control animals the highest histomorphological changes were observed on the 4th day after the infection in the liver, kidneys and spleen. In the animals with staphylococcal infection such changes were also observed in the lung tissue. In the animals treated with prodigiozan or S-methylmethionine in a dose of 20 mg or 0.3 ml of a 5 per cent solution respectively simultaneously with or 2 days after the infection the changes in the organs were less pronounced than those in the control. Under the effect of prednizolone used in a dose of 10 mg according to the same schedule the inflammatory-dystrophic changes in the internal organs of the animals with dysentery and staphylococcal infections were much higher as compared to the control.     

Mitogenic effect of Legionella pneumophila: the ratio of proliferating T- and B-cells changes after immunization

The splenic lymphocyte proliferative response of guinea-pigs immunized with L. pneumophila antigen (ALP) was studied. The immune animals were shown to express enhanced reactivity in response to E. coli lipopolysaccharide, as compared to the controls, while the response to concanavalin A was markedly decreased. After 3 days in the culture ALP induced a significant nonspecific lymphocyte transformation that was expressed to a similar degree in both experimental and control group. However, after a 5-day incubation ALP induced a specific proliferative response of the immunized guinea-pig splenocytes. The experiments on fractionation by passing through the column with nylon wool revealed that T cells are activated in non-immunized guinea-pigs. The results obtained prove that T cells mediate the immune response to ALP in guinea-pigs.     

脾动脉栓塞术后出现严重粘连的脾切除手术体会

目的:探讨脾动脉栓塞术后严重脾粘连脾切除手术技巧,为临床实践提供可借鉴的方法。方法:收集我院2005年4月-2013年6月收治的18例脾动脉栓塞术后严重脾粘连行脾切除术的患者临床资料,分析手术时间、术中出血、术中特殊处理及术后恢复情况等。结果:18例患者均顺利恢复出院,无围手术期死亡,开腹后到脾切除完成平均耗时55 min、出血550 mL。术后并发症为腹水(8例)、肺部感染(1例)、胰瘘(1例)及腹腔内出血(1例)。结论:脾动脉栓塞术后出现严重脾粘连行脾切除术,手术风险较大,手术时间、术中出血较常规脾切除术明显延长、增多。规范手术操作,细致分离脾周粘连,合理处理脾蒂,是安全、有效完成此类脾切除术的关键。     

Blastogenesis of splenic lymphocytes to specific antigens and PHA in Paragonimus westermani infected mice]

Paragonimus westermani is a common fluke in Korea. The present study aimed to observe the cell mediated immune response in experimental paragonimiasis of mice. The mouse (BALB/c) was orally inoculated with 40 metacercariae of P. westermani from Cambaroides similis. During the infection (1, 2, 4, 6 weeks) of mouse, blastogenic response of splenic lymphocytes to P. westermani adult antigen, metacercaria antigen, and PHA were observed. Sera from infected and noninfected mice added to normal mouse splenic lymphocytes with or without PHA. The blastogenic response of splenic lymphocytes to PHA was reduced after 1 week of infection. However after 6 weeks of infection, the response was restored to the control level. The blastogenic response of splenic lymphocytes to P. westermani adult or metacercaria antigen increased significantly on 1 week after infection, and maintained up to 6 weeks after infection. The response of non-infected mice was suppressed by addition of the infected mouse serum. The present results suggested that cellular immunity was involved in P. westermani infected mice and that P. westermani anti-serum inhibited proliferation of T lymphocytes.     

Macrophage-mediated suppression of con A-induced IL-2 production in spleen cells from syphilitic rabbits

Blast transformation studies have indicated a diminished T cell response in spleen cell preparations from rabbits infected with Treponema pallidum. IL-2 synthesis by T lymphocytes is required for proliferation of these cells. Thus, Con A-induced IL-2 generation was measured in syphilitic animals infected for 9 to 14 days. IL-2 production in the infected rabbits was only one-half that observed for uninfected rabbits. This marked decrease in IL-2 was not caused by decreased IL-1 secretion by adherent cells from infected animals because similar levels were found in both infected and uninfected splenic cultures. This decrease was also not caused by an increase in infected spleen cell adsorption of IL-2; similar numbers of receptors for this IL were present in Con A-stimulated infected and uninfected splenic preparations. The inhibited IL-2 production in infected spleen cells was reversed upon removal of the adherent cells and also elevated upon addition of indomethacin to the cultures. PGE levels were also elevated in splenic cultures from infected animals. Finally, IL-2 synthesis, when evaluated at various days postinfection, showed that at 4 days, splenic cells generated twice as much IL-2 as uninfected cells. At 9 to 14 days, IL-2 levels were dramatically decreased (50% lower than that observed in uninfected cultures), and suppression of IL-2 by adherent cells was observed as late as 35 days post-infection. We propose that premature down regulation (suppression) of IL-2 secretion is mediated by adherent cells via a cyclo-oxygenase product, most likely PGE. These results may explain why most, but not all, treponemes are cleared during infection, and why the secondary manifestations of the disease occur.     

Immune responses of DBA/2 mice bearing melanoma tumors: Cell-mediated immune responses after challenge with vaccinia virus

Summary Cell-mediated immune responses in DBA/2 mice bearing melanoma tumors (TB-mice) were measured and compared to similar responses in mice without tumors (C-mice). Splenic lymphocytes from TB-mice had a reduced capacity to respond to both B and T-cell mitogens, but TB-mice responded to infection with vaccinia virus by developing a virus-specific cytotoxic T-cell response equal to that measured with splenic effectors prepared from virus-infected C-mice. NK-cell activity, as measured by the in vitro lysis of YAC-1 targets by splenic effectors, was significantly depressed in TB-mice but, after infection of the animals with vaccinia virus, was restored to levels equal to that measured with splenic effectors prepared from C-mice. Doses of vaccinia virus, strain WR which elicited vaccinia-virus-specific cytotoxic T cells or stimulated NK-cell activity, failed to elicit or stimulate cytotoxic effectors specific for S91-melanoma tumor cells.     

Rotavirus-specific cytotoxic T lymphocytes appear at the intestinal mucosal surface after rotavirus infection.

The gastrointestinal tract is constantly exposed to a variety of potentially invasive bacteria and viruses. The first line of defense of the host against these pathogens is the intestinal mucosal surface, which consists of epithelial cells, intraepithelial lymphocytes (IELs), mucus, and secretory immunoglobulins. Little is known about the function, memory, or trafficking of IELs after intestinal infection. We found that IELs obtained 6 days after oral inoculation of mice with the intestinal pathogen rotavirus (simian strain RRV) lysed rotavirus-infected target cells; cytotoxic T lymphocytes (CTLs) were responsible for rotavirus-specific cytotoxic activity. Rotavirus-specific cytotoxic activity by IELs was (i) eliminated by treatment with Thy 1.2-specific immunoglobulin M plus complement, (ii) restricted by proteins encoded at the major histocompatibility complex, and (iii) absent in mock-infected animals. Oral inoculation of mice with RRV also induced rotavirus-specific CTLs in splenic and intestinal lymphocytes (mesenteric lymph nodes, Peyer's patch). Parenteral inoculation induced rotavirus-specific CTLs in splenic, intestinal (IELs, mesenteric lymph nodes, Peyer's patch), and nonintestinal lymphocytes (inguinal nodes). Therefore, presentation of rotavirus to the intestinal mucosal surface was not necessary to induce IELs with virus-specific cytotoxic activity. At 4 weeks after oral or parenteral inoculation of mice with RRV, rotavirus-specific CTL precursors appeared among splenic, Peyer's patch, inguinal, and mesenteric node lymphocytes, but not among IELs. IELs with rotavirus-specific cytotoxic activity may be generated from precursors at a site other than the intestinal mucosal surface. Part of the response of the host to enteric infection may include surveillance and lysis of virus-infected villus epithelial cells by IELs.     

Partial splenic embolisation for hypersplenism of thalassaemia major: five year follow up.

Six patients with thalassaemia major were treated by partial splenic embolisation as an alternative to splenectomy and followed up for five years. Results were compared with those in a matched control group of seven patients treated by splenectomy. All patients treated by partial splenic embolisation showed a reduction in blood transfusion requirements comparable with those in the controls and which remained unchanged over the five years. Serious infections that commonly occur in patients splenectomised for thalassaemia did not occur after embolisation, presumably owing to preservation of some immune function by the splenic remnant. By contrast with the change in platelet counts seen after splenectomy, platelet counts remained normal after partial splenic embolisation, so reducing the risk of thromboses. On the other hand, pre-existing leucopenia and thrombocytopenia were corrected after embolisation. It is concluded that partial splenic embolisation provides an alternative to splenectomy for thalassaemia major and is equally effective and much safer.     

急性巨细胞病毒感染导致新生豚鼠脾细胞亚群的定量改变

新生豚鼠皮下接种豚鼠巨细胞病毒(GPCMV)后,导致脾脏GPCMV急性感染,脾脏肿大,脾细胞增生,计算表明,脾T淋巴细胞,B淋巴细胞、吞噬细胞数显著高于正常动物,接种病毒后第5天即可在脾细胞检出高滴度感染性病毒(达2.5 log10 TCID50/10~7细胞),其中,主要是脾B淋巴细胞、T淋巴细胞和巨噬细胞携带病毒。     

Therapy of experimental staphylococcal infection with a hyperimmune antistaphylococcal plasma

Generalized nonlethal infection was induced in guinea pigs by the intramuscular injection of 10(10) S. aureus cells, and on days 1, 3 and 5 after inoculation a half of the animals received antistaphylococcal hyperimmune human plasma intraperitoneally in a dose of 2 ml (6 AU/ml) per animal. This plasma decreased the number of staphylococcal colonies in the spleen and affected mainly the system of phagocytes (neutrophils): on day 3 the percentage of neutrophils with Fc gamma R increased in the treated guinea pigs in comparison with the controls and with the initial level; from day 6 the percentage of active neutrophils with Fc gamma R increased in the treated animals in comparison with the controls; on day 14 the percentage of such neutrophils increased in comparison with the initial level as well, and the treated guinea pigs also showed an increase in the content of lymphocytes with receptors for staphylococci in comparison with the controls. There was little difference in the content of neutrophils with CR, as well as in the percentage of active phagocytes with these receptors, in the treated animals and in the controls. By the moment of the resolution of generalization (day 14) the content of active T-lymphocytes increased in the treated guinea pigs in comparison with the controls and with the initial level; simultaneously the levels of B-lymphocytes and B-lymphocytes with receptors for mouse red blood cells returned to their initial values.     

Thymic and splenic changes following injection of living Brucella and BCG in the Guinea pig

Summary The thymic and splenic reactions, following injections of BCG and living Brucella M. in the guinea pigs, were studied. Both microorganisms injected intravenously produced thymic involution, maximal after BCG inoculation, followed by regeneration that was complete by day 10 in Brucella M.-treated guinea pigs, and by day 15, in BCG injected guinea pigs. Increase of mitotic index was more accentuated and more persistent after Brucella M. than after BCG treatment in the thymus. After a short involution period the spleen of injected animals increased in size in both groups. The splenic enlargement was dramatic and occurred at an accelerated rate in animals given BCG. It appeared to be the result of a conspicuous involvement of the red pulp by multiple granulomas. In Brucella M. treated guinea pigs the splenic enlargement was less obvious, but the splenic white pulp was more abundant in BCG-treated guinea pigs. Granulomas were observed only in the periarterial sheaths of the white pulp.These observations provide evidence for the hypothesis that injections of both BCG and Brucella M. provoke a proliferation of B and T lymphocytes, a migration of T lymphocytes from the thymus to the T-dependent area of the spleen which seems, perhaps, more marked after injection of Brucella M., and a strong granulomatous histiocytic reaction which is more conspicuous in animals given BCG.     

Evidence of immunological activity in heterotropic autotransplanted splenic tissue in DBA/2 mice

After total splenectomy, a portion of the spleen was autotransplanted into subcutaneous ectopic sites of DBA/2 mice. These implanted fragments regenerated into splenic tissue that are microscopically indistinguishable from the structure of the original spleen. These implants are also immunologically functional as demonstrated by their ability to produce antibodies to sheep red blood cells (SRBC) and SRBC inhibition of splenic (implant) cell migration.     

Splenectomy associated changes in IgM memory B cells in an adult spleen registry cohort

Asplenic patients have a lifelong risk of overwhelming post-splenectomy infection and have been reported to have low numbers of peripheral blood IgM memory B cells. The clinical value of quantitation of memory B cells as an indicator of splenic abnormality or risk of infection has been unclear. To assess changes in B cell sub-populations after splenectomy we studied patients recruited to a spleen registry (n = 591). A subset of 209 adult asplenic or hyposplenic subjects, and normal controls (n = 140) were tested for IgM memory B cells. We also determined a) changes in IgM memory B cells with time after splenectomy using the cross-sectional data from patients on the registry and b) the kinetics of changes in haematological markers associated with splenectomy(n = 45). Total B cells in splenectomy patients did not differ from controls, but memory B cells, IgM memory B cells and switched B cells were significantly (p<0.001) reduced. The reduction was similar for different indications for splenectomy. Changes of asplenia in routine blood films including presence of Howell-Jolly bodies (HJB), occurred early (median 25 days) and splenectomy associated thrombocytosis and lymphocytosis peaked by 50 days. There was a more gradual decrease in IgM memory B cells reaching a stable level within 6 months after splenectomy. IgM memory B cells as proportion of B cells was the best discriminator between splenectomized patients and normal controls and at the optimal cut-off of 4.53, showed a true positive rate of 95% and false positive rate of 20%. In a survey of 152 registry patients stratified by IgM memory B cells around this cut-off there was no association with minor infections and no registry patients experienced OPSI during the study. Despite significant changes after splenectomy, conventional measures of IgM memory cells have limited clinical utility in this population.     

The function of the liver mitochondria in rats with experimental cirrhosis undergoing an organ-preserving operation on the spleen]

The effect of splenectomy and of spleen-preserving operation with suture ligation of the left gastric artery on the functional status of liver mitochondria was studied by using 102 white male-rats of mixed population with experimental cirrhosis. The obtained data made it evident that in the immediate postoperative period (from 1 to 3 weeks after the procedure) in the animals of both the first and the second series, the disorder of energetic regulation of mitochondria hepatocytes respiration and the decrease in phosphorylation efficiency had the same tendency, which were seemingly caused by stress-action of the operative trauma. The data accumulated in the more distant postoperative period (8 weeks after the procedure) indicated that the resection of the lower splenic pole in combination with supplementary liver arterialization improved essentially the functional status of mitochondria hepatocytes and was more beneficial in contradistinction to splenectomy.     

Lymphocyte plasma membranes. III. Composition of lymphocyte plasma membranes from normal and immunized rats

Isolated plasma membranes of thymic and splenic lymphocytes from unimmunized and immunized rats of the inbred ACI and F344 strains were analyzed for chemical and enzymatic composition, for membrane protein patterns by polyacrylamide gel electrophoresis and for membrane-associated immunoglobulins. After immunization, the thymic and splenic lymphocyte membranes from F344 rat contained less carbohydrate and higher phospholipid contents than control animals. In both ACI and F344 inbred rat strains the membrane phospholipid to cholesterol weight ratio increased significantly after immunization. The electrophoretic patterns of solubilized membrane proteins and of iodinated external membrane proteins were similar in unimmunized and immunized animals.When thymic and splenic lymphocytes of normal or immunized animals were surface radioiodinated, solubilized in Triton X-100, NP-40 or 10 M urea in 1.5 M acetic acid and analyzed by immunoprecipitation, labeled IgM immunoglobulin was recovered from thymic lymphocytes but both labeled IgG and IgM were recovered from splenic lymphocytes. However, when unlabeled isolated plasma membranes were solubilized in 1% Triton X-100 and analyzed by immunodiffusion in agarose gels, both IgG and IgM were identified in thymic and splenic cells.     

Dynamics of staphylococcal infection in guinea pigs with delayed hypersensitivity to Staphylococcus aureus surface antigens

The relationship between delayed hypersensitivity (DH) to S. aureus surface antigens and the intensity of the infectious process induced by the sublethal infection of guinea pigs with S. aureus was studied. The protective effect, manifested by a decrease in the staphylococcal contamination of the spleen tissue and by an increase in the level of the activation of lymphocytes, was shown to correlate with DH induced by inactivated staphylococcal cells. In infected guinea pigs having DH to different staphylococcal antigens the disease either took a more severe course (in cases of DH to cell wall or peptidoglycan) than in the animals subjected only to infection, or no aggravation of the disease was observed (in cases of DH to protein A).