Regulation of the growth and differentiation of Trypanosoma (Trypanozoon) brucei brucei in resistant (C57B1/6) and susceptible (C3H/He) mice

While Trypanosoma brucei brucei GUTat 3 were equally infective for C3H/He and for C57Bl/6 mice at doses ranging from 5 to 5 x 10(3) organisms and had similar prepatent periods in both strains of mice, infected C57Bl/6 mice displayed lower parasitaemia, shorter times to parasite wave remission and survived for a longer time than infected C3H/He mice. Parasite growth and differentiation rates and host immune responses were similar for the first 5 days in both strains of mice after infection with 10(3) T.b.brucei GUTat 3 but, thereafter, parasite differentiation proceeded more rapidly and specific antibodies reached higher titres in C57Bl/6 than in C3H/He mice. In contrast, parasite growth and differentiation rates were similar in irradiated mice of both strains. Furthermore, following inoculation of intact mice with irradiated T.b.brucei GUTat 3, C3H/He mice actually mounted higher titred antibody responses than C57Bl/6 mice showing that they were not intrinsically defective in their capacity to respond to GUTat 3 antigens. Parasite differentiation occurred at the same rate in irradiated (650r) C57Bl/6 mice and in irradiated C57Bl/6 mice reconstituted with syngeneic spleen cells although T.b.brucei GUTat 3 specific antibody was detected in the latter mice prior to peak parasitaemia. Furthermore, it was shown directly in C57Bl/6 mice that there was no selective destruction of slender form T.b.brucei GUTat 3 parasites during the phase of accumulation of stumpy form parasites. These studies indicate that the more rapid differentiation of T.b.brucei GUTat 3 parasites in infected C57Bl/6 mice as compared to infected C3H/He mice was unlikely to be directly related to the more efficient antibody response in the infected C57Bl/6 mice. The observations suggest that there might be an association between host mechanisms which regulate differentiation of T.b.brucei parasites and those which regulate antibody responses.     

Regulation of parasite-specific antibody responses in resistant (C57BL/6) and susceptible (C3H/HE) mice infected with Trypanosoma (trypanozoon) brucei brucei

After infection with 10(3) T. brucei GUTat 3.1, C57BL/6 mice produced antibody responses and controlled the first parasitaemic wave whereas C3H/He mice did not. The inability of C3H/He mice to control parasitaemia resulted from an impaired ability of parasite-induced antibody-containing cells to secrete immunoglobulin. Antibody-containing cells in infected C3H/He mice regained the ability to secrete antibody within 24 h after trypanosome elimination by treatment with Berenil, suggesting that the block in antibody secretion was maintained by living parasites or short-lived components of degenerating parasites. Infected C3H/He mice also had an impaired ability to produce a rabbit erythrocyte-specific antibody response on challenge with rabbit erythrocytes and this response recovered when parasites were eliminated from the blood 24 h before analysis. It was not possible to inhibit secretion of antibody by rabbit erythrocyte-induced plasma cells either by incubating them with serum from infected C3H/He mice or by injecting large numbers of living trypanosomes into C3H/He mice already responding to rabbit erythrocytes. The process leading to failure of parasite and rabbit erythrocyte-induced antibody-containing cells to become high rate antibody-secreting cells was not identified but did not appear to correlate with any obvious change in the intra-cellular morphology of the antibody-containing cells.     

Analysis of B cell and T cell proliferative responses induced by monomorphic and pleomorphic Trypanosoma brucei parasites in mice

Cells collected from C57B1/6 mice infected with monomorphic and pleomorphic clones of Trypanosoma brucei parasites (ILTat 1.4 and GUTat 3.1) were analysed for the incorporation of 125I-Iododeoxyuridine into DNA of total splenic lymphocytes and of B and T lymphocytes isolated on a fluorescence activated cell sorter. The monomorphic T. brucei ILTat 1.4 parasites triggered delayed and low splenic DNA synthetic responses in comparison to those arising in mice infected with the pleomorphic T. brucei GUTat 3.1 organisms. Mice infected with both parasite clones mounted splenic DNA synthetic responses similar to those arising in animals infected with the pleomorphic organisms alone and similar responses were induced by lethally irradiated T. bruceiGUTat 3.1 and T. brucei ILTat 1.4 parasites. In mice infected with the pleomorphic parasites, DNA synthesis was first detected in the T cell population and B cell DNA synthetic responses were detected between 1 and 2 days later. In contrast only T cell DNA synthetic responses were detected after infection with the monomorphic T. brucei ILTat 1.4 parasites. It is suggested that the previously reported failure of monomorphic T. brucei parasites to trigger antibody production in infected mice is a result of their inability to stimulate B lymphocytes.     

Trypanosoma brucei infection in nude mice: B lymphocyte function is suppressed in the absence of T lymphocytes

B lymphocyte function was assessed in outbred nude mice and nu/+controls infected with Trypanosoma brucei brucei. On day 10 of the infection in outbred nu/nu mice in which the initial wave of parasites was strongly controlled, B cell function was unaltered on enhanced compared with uninfected animals or infected nu/+. In other nu/nu mice unable to control the initial parasitaemia, thymidine incorporation and Ig secretion by spleen cells were increased on day 10 and their response to lipopolysaccharide in vitro negated. By day 15 however, even the spleen cells of infected nu/nu which controlled the initial wave of parasites were proliferating and secreting Ig on removal from the mice and they were unable to respond to LPS in vitro. These experiments confirm results of a previous study of B cell function in T cell-depleted mice (Askonas et al. 1979). T. b. brucei infection of mice causes both enhanced Ig production and suppression of the ability of B cells to respond to mitogen even in the absence of T cells, but the presence of T cells may accelerate the changes which occur in B lymphocytes following this infection.     

Trypanosoma brucei variable surface antigen is released by degenerating parasites but not by actively dividing parasites

Surface antigen biosynthesis and fate in monomorphic and pleomorphic Trypanosoma brucei was examined to assess how slender and stumpy form T. brucei parasites present their variant specific glycoprotein (VSG) to the host immune system. Monomorphic and pleomorphic T. brucei did not release recently synthesized VSG in vitro. Slender form T. brucei , either from monomorphic or pleomorphic populations, did not release VSG in vivo. Detection of free VSG in plasma from irradiated mice infected with pleomorphic parasites correlated with the appearance of stumpy form parasites and possibly arose as a result of degeneration of those parasites. The in vivo released VSG was found to react well with some but not all antibodies directed against VSG determinants. Monoclonal and monospecific antibodies which react with VSG on living trypanosomes did not react with the released VSG whereas VSG-specific monoclonal antibodies which do not react with the surface of living T. brucei did react with the released VSG. It was unclear whether released VSG had lost a conformational determinant expressed on trypanosome-attached VSG or whether antibodies which react strongly with VSG on living trypanosomes are of such low avidity that they fail to bind released VSG. The results suggest that trypanosome-attached VSG is more important for stimulation of protective humoral responses than released VSG. The requirements for stimulation of protective anti-VSG responses are reported elsewhere (Sendashonga & Black 1982).     

Cytokines and antibody responses during Trypanosoma congolense infections in two inbred mouse strains that differ in resistance

We studied IL-4, IL-10 and IFN-gamma secretion by splenocytes and the plasma levels of different isotypes of antibodies against various antigens of Trypanosoma congolense in highly susceptible BALB/c and relatively resistant C57BL/6 mice during the early course of infection with T. congolense. The patterns of appearance of cytokine spotforming cells in the spleens were essentially similar in the two mouse strains although higher numbers were detected in the spleens of BALB/c than C57BL/6 mice on some days post-infection. However, the amount of IL-4, IL-10 and IFN-gamma secreted into the culture fluids was dramatically different. From day 4 forward, splenocytes from BALB/c mice secreted very high levels of these cytokines. In contrast, splenocytes from infected C57BL/6 mice did not secrete detectable levels of IL-4 throughout the period tested. The secretion of IL-10 and IFN-gamma by C57BL/6 splenocytes only became appreciable on day 6 and was down-regulated by day 8, when the first wave of parasitaemia was being controlled. At days 6-8, splenocytes from infected C57BL/6 mice secreted two-fold higher amounts of IL-12 p40 than those from BALB/c mice. Infected BALB/c mice mounted an earlier IgM antibody response to variant surface glycoprotein (VSG), formalin-fixed T. congolense and whole T. congolense lysates than did infected C57BL/6 mice. However, they failed to make any detectable IgG3 and IgG2a antibody responses to these antigens whereas infected C57BL/6 mice made strong IgG3 and IgG2a responses. We speculate that enhanced resistance against T. congolense infections in mice may be mediated by IL-12 dependent synthesis of IgG2 antibodies to VSG and possibly also common trypanosomal antigens.     

不同品系小鼠对卡氏肺孢子虫的易感性及其免疫反应的研究

目的　探讨不同品系小鼠对卡氏肺孢子虫 (P.c)的易感性及其免疫反应。　方法　以接触传播方式使BAL B/ c和 C5 7BL / 6小鼠 (各 15只 )感染 P.c。观察小鼠与传染源接触 4、5、6 wk后肺内虫数、支气管肺泡灌洗液中CD4 + T细胞和 CD8+ T细胞及血清 Ig G水平的变化。　结果　与传染源接触 4 wk,小鼠肺内均可检出 P.c,之后 ,BAL B/ c小鼠虫数继续增高 ,5 wk末达高峰。4 wk时 C5 7BL/ 6小鼠虫数无明显变化 ,5 wk时显著少于 BAL B/ c小鼠。两组小鼠支气管肺泡灌洗液内 CD4 + CD6 2 low T细胞和 CD8+ CD6 2 low T细胞数明显增多 ,血清 Ig G抗体水平显著上升。6 wk小鼠肺内 P.c均转阴。　结论　 BAL B/ c比 C5 7BL / 6小鼠更易感染 P.c。 BAL B/ c和 C5 7BL / 6小鼠感染 P.c后5 wk可产生有效的细胞和体液免疫反应并清除肺内感染的 P.c     

Suppressive macrophages occurring in murine Trypanosoma brucei infection inhibit T-cell responses in vivo and in vitro

Intraperitoneal injection of Trypanosoma brucei AnTat 1.1 into mice of the C3H.He, BALB/c or C57BL/6 strains resulted in impaired immune responses from day 3 onwards, as measured by the reduction in DNA synthesis in spleen cell populations stimulated with concanavalin A (Con-A) in vitro. Adherent cells from the peritoneum (PC) or from the spleen of infected mice, consisting predominantly of macrophages, caused a 60-80% reduction of the Con-A response in spleen cells from syngeneic recipients 3-4 days after transfer in vivo. Adherent PC from irradiated or athymic mice were equally suppressive. Spleen cells from infected mice reduced the proliferative response of spleen cells from uninfected mice upon co-cultivation in vitro. This dominant suppressive effect was abolished after the selective removal of macrophages from the spleen cell population by treatment with L-leucine methylester. Moreover, the macrophage-depleted spleen cells from infected mice responded normally to Con-A provided they were supplemented with splenic adherent cells from naive mice as a source of accessory cells. Both the cell transfer and co-cultivation experiments suggest that infection with African trypanosomes changes the properties of macrophages to a state which allows them actively to suppress immune responses.     

Immunoregulation in experimental murine Trypanosoma congolense infection: anti-IL-10 antibodies reverse trypanosome-mediated suppression of lymphocyte proliferation in vitro and moderately prolong the lifespan of genetically susceptible BALB/c mice

We infected highly susceptible BALB/c and relatively resistant C57BL/6 mice with cloned Trypanosoma congolense and followed the effects of these infections on the circulating parasite numbers, mouse mortality and cytokine expression. C57BL/6 mice controlled their parasitaemia and survived for up to 163 ± 12 days, while BALB/c mice could not control their parasitaemia and succumbed to the infection within 8.4 ± 0.5 days. Susceptible BALB/c mice had dramatically higher plasma levels of IL-10 than the resistant C57BL/6 mice from day 7 forward. This was preceded by an earlier and higher level induction of splenic IL-10 messenger RNA (mRNA) expression in the infected BALB/c mice. There was a strong negative correlation between the splenocyte proliferative responses to Concanavalin-A (Con-A) and their production of IL-10 in these infected BALB/c mice. Co-treatment of the Con-A-stimulated spleen cell cultures with monoclonal anti-IL-10 antibodies, but not isotype-matched control antibodies, could completely reverse this suppression of the splenocyte proliferative response. Finally, in three experiments, anti-IL-10 antibody treatment in vivo reduced the peak circulating parasitaemia of infected BALB/c mice by 43% and increased their median survival periods by 38% relative to isotype-matched control antibody-treated mice .     

Humoral responses against Trypanosoma brucei variable surface antigen are induced by degenerating parasites

An analysis was made of the inductive stimuli for anti-T. brucei variant surface glycoprotein (VSG) responses and the role played by humoral immunity in trypanosome wave control. The first wave parasitaemia was influenced by the rate of parasite differentiation from rapidly dividing slender forms to short lived stumpy forms. Remission of first wave parasitaemia was caused by a humoral immune response against external determinants of surface expressed VSG. Anti-VSG responses were accompanied by anti-trypanosome plasma membrane responses and were followed by non-specific responses. Responses appeared to be initiated by fragments of parasites on which VSG external determinants and plasma membrane antigens were accessible and were possibly accelerated and amplified by a trypanosome mitogen which was not VSG. The parasite fragments may have arisen as a result of degeneration of stumpy form but not slender form parasites.     

Secondary bacterial infection in plasma endotoxin levels and the acute-phase response of mice infected with Trypanosoma brucei brucei

Murine Trypanosoma brucei brucei infection leads to elevated plasma endotoxin-like activity levels not related to parasitaemia levels accompanied by the development of acute-phase response and increased plasma levels of serum amyloid P (SAP) and haptoglobin (Hp). To determine the source of the endotoxin-like activity and role of secondary bacterial infection in the pathogenesis of trypanosomosis, infected mice were treated with the antibiotic ciprofloxacin. Plasma endotoxin-like activity levels, irrespective of treatment, were elevated three- to fourfold, beginning 7 days after infection. Plasma protein concentrations increased markedly following infection from 7 days after infection (DAI). Peak Hp and SAP concentrations in ciprofloxacin-treated and -untreated infected mice were attained 7 and 14 DAI, respectively. Thereafter, both protein levels gradually declined until the end of the experiment, but Hp levels for non-treated mice declined up to 21 DAI and thereafter significantly increased on 28 and 35 DAI. Whole-trypanosome lysate and the membrane-enriched fraction demonstrated endotoxin-like activity, with the former having higher levels. The results suggest that the endotoxin-like activity in trypanosome fractions and plasma of infected mice is due to the trypanosome. Further elevation of haptoglobin during the late stages of infection in non-treated mice suggests the involvement of secondary bacterial infection.     

Factors affecting the infection of the D variant of encephalomyocarditis virus in the B cells of C57BL/6J mice

Summary The D variant of encephalomyocarditis viras is capable of infecting most inbred strains of mice. However, only certain strains are susceptible to the diabetogenic effect of this viras. In order to understand why some inbred strains do not become diabetic, the pathogenesis of infection was studied in diabetes-resistant C57BL/6J mice. It was the purpose of the investigation to ascertain whether specific host defense factors might play a crucial role in the mechanism of resistance. To determine whether perturbations of the immune response would alter the resistance of these animals, mice were treated with a high dose (1.15 mmol/kg body weight) of the T- and B-cell toxin cyclophosphamide prior to infection with the D variant. This treatment did not induce overt diabetes or glucose intolerance in the mice tested 7 days after infection. Based on this finding, it appeared likely that resistance to the D variant is conveyed by some factor other than cell-mediated immunity. A likely candidate to control this viral infection is the interferon system. To investigate this possibility, C57BL/6J mice were infected with the D variant and the concentrations of serum interferon titred at various intervals thereafter. In contrast to previous reports with diabetes susceptible mice, C57BL/6J mice were found to generate a substantial interferon response against this variant, with peak levels found in the serum at 24 h following infection. Additional studies were performed in which mice were treated with antibody to mouse interferon alpha/beta at the time of infection and again 3 days after infection with the D variant. Sixty percent of the animals treated in this manner became glucose intolerant. The results of these studies indicate that the interferon system is a critical determinant of resistance to the diabetogenic effect of the D variant in C57BL/6J mice.     

Susceptibility of severe combined immuno-deficient (SCID) mice to Trypanosoma brucei gambiense and T. b. rhodesiense

Susceptibility of severe combined immunodeficient (SCID) mice to 7 isolates of Trypanosoma brucei gambiense and 2 isolates of T. b. rhodesiense was examined in terms of their infectivity, course of parasitaemia, packed cell volume (PCV) and survival period in comparison with that of normal immunocompetent (BALB/c) mice. All isolates of T. b. gambiense and T. b. rhodesiense caused high (> 1 × 10⁸ parasites/ml) parasitaemia in the SCID mice, the survival periods ranged from 5 to 47 days. On the other hand, 5 of 7 isolates of T. b. gambiense developed chronic infection in the BALB/c mice with sporadic but persistent parasitaemia with less than 5 × 10⁶ parasites/ml. All the mice tested in this group survived more than 60 days after infection. In contrast, the 2 remaining isolates of T. b. gambiense and both isolates of T. b. rhodesiense showed high virulence in the BALB/c mice and killed all of them within 30 days after infection. The results demonstrate that the SCID mice, in which functional B- and T-cell-mediated immunities are congenitally lacking, are highly susceptible for 'low-virulence' T. b. gambiense . This makes SCID mice useful tools for the isolation of parasites from T. b. gambiense sleeping sickness patients and the propagation of large amounts of such parasites.     

Control of hemopoiesis in mice by sensitized L3T4+ Lyt2-lymphocytes during infection with bacillus Calmette-Guérin.

When injected intravenously with bacillus Calmette-Guérin (BCG; 10(7) viable units), C57BL/6 mice rapidly develop a transient anemia associated with an increased number of granulocytes and monocytes, whereas C3H/He mice do not. Because these two features are lacking in C57BL/6 nude mice we postulated that T lymphocytes can regulate hemopoiesis during infection. To assess further the role in hemopoiesis of T lymphocytes present in bone marrow of C57BL/6 and C3H/He mice, the frequency of BCG-specific T lymphocytes and their surface marker phenotype were determined by limiting dilution analysis and use of monoclonal antibodies. The number of BCG-specific T lymphocytes was estimated to be 50- to 100-fold higher in bone marrow of C57BL/6 than in that of C3H/He mice. Although L3T4+ Lyt2-and L3T4- Lyt2+ BCG-specific T lymphocytes were generated in mice of both strains, in C57BL/6 mice L3T4+ cells were induced preferentially from day 1 through day 5 after infection in correlation with hemopoietic changes. The relation between T-cell immune response and hemopoietic changes was substantiated by results obtained after in vivo treatment with monoclonal antibodies. Selective depletion of L3T4+ T cells by in vivo injection of anti-L3T4 monoclonal antibodies (GK 1-5) inhibited the development of the anemia and the related increased production of phagocytes in C57BL/6 mice receiving BCG.     

Murine tumour necrosis factor plays a protective role during the initial phase of the experimental infection with Trypanosoma brucei brucei

Soluble extracts from salivarian trypanosomes (Trypanosoma brucei brucei, T. evansi and T. congolense) were shown to be capable of inducing murine tumour necrosis factor (mTNF) secretion, both in vivo and in vitro, whereas the soluble extract of an intracellular trypanosome (T. cruzi) failed to do so. Furthermore, the role of mTNF during the initial phase of experimental infections with T. brucei was studied by treating infected mice with mTNF-inducing trypanosoma soluble extract and with neutralizing monoclonal anti-mTNF antibodies. Treatment of the infected animals with different doses of T. brucei soluble extract resulted in a lower first parasitaemia peak (low lysate dose) and in a longer survival time or in a nearly total inhibition of parasite development (high lysate dose). Cotreatment of the infected mice with both anti-mTNF antibodies and a high dose of soluble extract completely restored the parasite development in both trypanosusceptible C3H/He mice and trypanosubtolerant CBA/Ca mice, indicating a protective role of mTNF during the parasitaemia. Collectively these results suggest a negative influence of mTNF on T. brucei development in vivo.     

Immune mechanisms in C57B1 mice genetically resistant to Trypanosoma congolense infection. II. Aspects of the humoral response

Some aspects of the humoral response in trypanotolerant C57B1 mice and susceptible A/J mice were investigated to determine the possible basis of trypanotolerance. When the hepatic uptake of 75Se-labelled T. congolense by infected mice was measured as an index of antibody production, it was found that only C57B1 mice could remove circulating labelled parasites, this ability persisting for several weeks after infection. Estimation of the immunoglobulin concentrations in both strains of mice showed that C57B1 mice developed a pronounced IgM response during the first parasitaemic wave, while A/J mice did not. Over the same period the IgM concentrations in C57B1 mice initially fell, but recovered at the time of peak parasitaemia. In contrast, A/J mice showed a continual fall in total IgG concentrations in the circulation until death 10 days after infection. Finally, it was shown that during the initial rising parasitaemia, the plaque forming cell responses of both strains of mice to sheep red blood cells were normal indicating that neither strain of mice was immunosuppressed. Also, A/J mice vaccinated with irradiated T. brucei on day 4 and C57B1 mice vaccinated either on day 4 or day 60 of a T. congolense infection were able to mount an effective immune response to the vaccine, as judged by the hepatic uptake of radiolabelled parasites. All of the results indicate that the trypanotolerance of C57B1 mice depends, at least in part, on their more efficient antibody response.     

Cerebral trypanosomiasis in cattle with mixed Trypanosoma congolense and T. brucei brucei infections

Six Boran steers were infected simultaneously with Trypanosoma congolense and T. brucei brucei while another group of 3 was inoculated with T. b. brucei one year after infection with T. congolense. Three further steers were infected with T. b. brucei alone. Whereas, the six animals which received simultaneous infections developed clinical signs of cerebral trypanosomiasis as evidenced by depression, ataxia and occasional circling, those infected with T. b. brucei alone did not. At necropsy, 4 out of the 6 simultaneously infected animals had a mild to severe disseminated non-suppurative meningoencephalitis. Trypanosoma b. brucei was isolated from the cerebrospinal fluid (CSF) of three out of the four animals with histological lesions. Two of the cattle superinfected with T. b. brucei one year after infection with T. congolense also developed both clinical and histological evidence of cerebral trypanosomiasis. Trypanosoma congolense was isolated from the CSF of one of these 2 animals. Specific antibodies to the variable surface glycoproteins (VSGs) of the infecting T. b. brucei and T. congolense clones were found in the CSF of the 8 animals that developed cerebral trypanosomiasis. In these animals however, there was neither temporal nor quantitative correlation between VSG-specific antibodies in serum and in CSF, implying a de novo synthesis of antibodies to the infecting trypanosomes in the CSF.     

Chemotherapeutic effects of Annona senegalensis in Trypanosoma brucei brucei

Mice infected with Trypanosoma brucei brucei 8/18 strain were treated orally and intramuscularly (im) with aqueous root extracts of Annona senegalensis, in doses of 27.8 mg kg-1 and 9.5 mg kg-1 respectively, for four consecutive days commencing 72 hours after the mice were infected. At these dosages the parasites were cleared from the circulation and no relapse was recorded over 60 days. The plant extract, however, had no effect on the trypanosomes when therapy was initiated at the late stages of infection, that is, about the sixth day when the parasitaemia level was 0.9 x 10(6); and all the animals died a day or two later. The herbal extracts also did not show any prophylactic action when given prior to infection. The root extract possesses different margins of safety in the mice depending on the route of administration. The therapeutic index for oral administration was 5.13, and that for im administration was 1.8. Chemical tests revealed that the plant extract contains alkaloids, saponins and tannins. Adverse reactions, especially to doses of 2.3-5.76 mg kg-1, were noted in animals that received the drug parenterally, but not when the drug was administered orally. However, A. senegalensis is shown to be therapeutically effective against T. b. brucei in mice, which agrees with the claims of Nigerian practitioners of Traditional Medicine that it is effective against trypanosomiasis in man.     

CTLA-4 regulates the murine immune response to Trypanosoma cruzi infection

Infection with Trypanosoma cruzi causes a profound suppression of T cell responsiveness to polyclonal or antigenic stimuli. In this study, we quantified expression of the negative T cell regulatory molecule CTLA-4 in T. cruzi infected mice and analysed its influence on the immune suppression. Levels of splenic CTLA-4 expression were highest around day 10 after infection, reaching 5% in resistant B6D2F1 mice, but exceeding 10% of CD4(+) T cells in C57BL/6 mice that were susceptible to mortal disease. The proliferative response of explanted splenocytes to CD3-mediated stimulation was strongly suppressed in both the susceptible and the resistant strains. Blockade of CTLA-4 in vitro with a monoclonal antibody affected neither proliferative response nor cytokine production (IFN-gamma, IL-4 and IL-2) by splenic T cells from infected C57BL/6 mice. Treatment of mice with anti-CTLA-4 antibody on the day of infection decreased IFN-gamma production and reduced mortality by about 50%. We conclude that high CTLA-4 expression is a hallmark of severe disease in murine T. cruzi infection, and that CTLA-4 has a regulative influence at the early stages during priming of the immune reaction to the parasite, augmenting a strong Th1-biased response.     

Trypanosoma brucei gambiense: cerebral immunopathology in mice

Ninety outbred white adult female mice were infected with Trypanosoma brucei gambiense (GUMS 2, alias LUMP 1237) originating from a Zairian patient and known to produce a low parasitaemia in rodents. The development of cerebral trypanosomiasis was independent upon the number of parasites inoculated per mouse. Trypanosomes appeared in the circulating blood about four months after infection, when some mice started to show the first signs of paresis which subsequently led to cachexia. A clinical test to stage such a development is described. 57 mice were sacrificed at various intervals after infection, starting from one to 22 months. The morphological changes in the brain consisted of a diffuse meningoencephalitis in 45 mice, (78.9%) often associated with parasites, the latter being best visualised in 21 mice (36.8%) by immunofluorescence using a specific antitrypanosome antibody. The trypanosomes showed a predominantly extravascular distribution in the cerebral parenchyma, to a lesser extent in the meninges and only rarely in the choroid plexuses. Deposits of immunoglobulins in the choroid plexuses and cerebral infiltrations by plasma cells were mild. The level of circulating immune complexes was found to be increased. Adequate intravenous Melarsoprol did not prevent the disease from progressing to advanced stages, and there is limited morphological evidence that it did not eradicate the parasite from the host. The immunofluorescent use of an antitrypanosome antibody to demonstrate the persistence of tissue parasites after chemotherapy is recommended. Murine models seem therefore to be suitable for drug screening in cerebral trypanosomiasis since all three trypanosomes of the brucei group can be adapted to mice.