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1.
Proc Natl Acad Sci U S A ; 98(24): 13984-9, 2001 Nov 20.
Article in English | MEDLINE | ID: mdl-11707582

ABSTRACT

Chlamydia trachomatis is an obligate intracellular human bacterial pathogen that infects epithelial cells of the eye and genital tract. Infection can result in trachoma, the leading cause of preventable blindness worldwide, and sexually transmitted diseases. A common feature of infection is a chronic damaging inflammatory response for which the molecular pathogenesis is not understood. It has been proposed that chlamydiae have a cytotoxic activity that contributes to this pathology, but a toxin has not been identified. The C. trachomatis genome contains genes that encode proteins with significant homology to large clostridial cytotoxins. Here we show that C. trachomatis makes a replication-independent cytotoxic activity that produces morphological and cytoskeletal changes in epithelial cells that are indistinguishable from those mediated by clostridial toxin B. A mouse chlamydial strain that encodes a full-length cytotoxin caused pronounced cytotoxicity, as did a human strain that has a shorter ORF with homology to only the enzymatically active site of clostridial toxin B. Cytotoxin gene transcripts were detected in chlamydiae-infected cells, and a protein with the expected molecular mass was present in lysates of infected epithelial cells. The protein was present transiently in infected cells during the period of cytotoxicity. Together, these data provide compelling evidence for a chlamydial cytotoxin for epithelial cells and imply that the cytotoxin is present in the elementary body and delivered to host cells very early during infection. We hypothesize that the cytotoxin is a virulence factor that contributes to the pathogenesis of C. trachomatis diseases.


Subject(s)
Bacterial Toxins/genetics , Chlamydia trachomatis/pathogenicity , Cytotoxins/genetics , Genes, Bacterial/physiology , Amino Acid Sequence , Bacterial Toxins/metabolism , Chlamydia trachomatis/genetics , Chlamydia trachomatis/growth & development , Cytotoxins/metabolism , Gene Expression , HeLa Cells , Humans , Molecular Sequence Data , Open Reading Frames
2.
Gene ; 275(2): 287-98, 2001 Sep 19.
Article in English | MEDLINE | ID: mdl-11587856

ABSTRACT

Cowdria ruminantium causes the tick-borne rickettsial disease of heartwater, which is devastating to livestock production in sub-Saharan Africa. Current diagnosis and control methods are inadequate. We have identified and sequenced a subset of genes encoding recombinant antigens recognized by antibody and peripheral blood mononuclear cells from immune ruminants. The identified genes include many with significant similarity to those of Rickettsia prowazekii, genes predicted to encode different outer membrane proteins and lipoproteins and a gene containing an unusual tandem repeat structure. Evidence is presented for immune protection by recombinant antigens in a mouse model of C. ruminantium infection. These data identify new recombinant antigens for evaluation in vaccines and diagnostic tests to control heartwater.


Subject(s)
Ehrlichia ruminantium/genetics , Genes, Bacterial/genetics , Immune System/immunology , Animals , Bacterial Proteins/genetics , Bacterial Proteins/immunology , Bacterial Vaccines/immunology , Cattle , Cell Division/immunology , Cell-Free System/metabolism , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Bacterial/isolation & purification , Ehrlichia ruminantium/immunology , Heartwater Disease/immunology , Heartwater Disease/microbiology , Heartwater Disease/mortality , Immune Sera/immunology , Immune System/microbiology , Lymphocytes/cytology , Lymphocytes/immunology , Lymphocytes/microbiology , Mice , Molecular Sequence Data , Protein Biosynthesis , Sequence Analysis, DNA , Sheep , Survival Rate , Transcription, Genetic
3.
Infect Immun ; 68(1): 192-6, 2000 Jan.
Article in English | MEDLINE | ID: mdl-10603387

ABSTRACT

Chlamydia trachomatis is a major cause of sexually transmitted disease (STD) for which a vaccine is needed. CD4(+) T-helper type 1 (Th1) cell-mediated immunity is an important component of protective immunity against murine chlamydial genital infection. Conventional vaccine approaches have not proven effective in eliciting chlamydial-specific CD4 Th1 immunity at the genital mucosa. Thus, it is possible that the development of a highly efficacious vaccine against genital infection will depend on the generation of a live attenuated C. trachomatis vaccine. Attenuated strains of C. trachomatis do not exist, so their potential utility as vaccines cannot be tested in animal models of infection. We have developed a surrogate model to study the effect of chlamydial attenuation on infection and immunity of the female genital tract by treating mice with a subchlamydiacidal concentration of oxytetracycline following vaginal infection. Compared to untreated control mice, antibiotic-treated mice shed significantly fewer infectious organisms (3 log(10)) from the cervico-vagina, produced a minimal inflammatory response in urogenital tissue, and did not experience infection-related sequelae. Antibiotic-treated mice generated levels of chlamydia-specific antibody and cell-mediated immunity equivalent to those of control mice. Importantly, antibiotic-treated mice were found to be as immune as control untreated mice when rechallenged vaginally. These findings demonstrate that subclinical chlamydial infection of the murine female genital tract is sufficient to stimulate a potent protective immune response. They also present indirect evidence supporting the possible use of live attenuated chlamydial organisms in the development of vaccines against chlamydial STDs.


Subject(s)
Chlamydia Infections/immunology , Chlamydia trachomatis/immunology , Genital Diseases, Female/immunology , Animals , Anti-Bacterial Agents/pharmacology , Antibodies, Bacterial/biosynthesis , Antibodies, Bacterial/blood , Antibody Specificity , Bacterial Vaccines/pharmacology , Chlamydia Infections/drug therapy , Chlamydia Infections/prevention & control , Chlamydia trachomatis/isolation & purification , Female , Genital Diseases, Female/drug therapy , Genital Diseases, Female/prevention & control , Interferon-gamma/biosynthesis , Interleukin-10/biosynthesis , Mice , Mice, Inbred C57BL , Oxytetracycline/pharmacology , Vaccines, Attenuated/pharmacology
4.
J Infect Dis ; 180(4): 1252-8, 1999 Oct.
Article in English | MEDLINE | ID: mdl-10479155

ABSTRACT

Chlamydia trachomatis is a major cause of sexually transmitted disease (STD) worldwide. Antibiotics are effective in treating infection; however, reinfection is common. This observation has led to the conclusion that infection fails to elicit a protective antichlamydial immune response. It was postulated that high reinfection rates might be due to early eradication of organisms from genital tissue after antibiotic intervention, which could negatively influence the development of naturally acquired protective immunity. This hypothesis was tested by use of a murine model of female genital infection. The findings show that doxycycline intervention of infection, although very effective in eradicating chlamydiae from genital tissue and preventing upper genital tract disease, significantly inhibits the development of protective immunity. If antibiotic intervention of human chlamydial genital infection has a similar effect on protective immunity, it could have important implications in the understanding of immunity to infection and future public health efforts to control chlamydial STD.


Subject(s)
Chlamydia Infections/drug therapy , Chlamydia Infections/immunology , Chlamydia trachomatis , Doxycycline/therapeutic use , Immunity, Innate/drug effects , Animals , Anti-Bacterial Agents/therapeutic use , Chlamydia Infections/prevention & control , Chlamydia trachomatis/immunology , Female , HeLa Cells , Humans , Mice , Mice, Inbred C57BL , Recurrence
5.
J Immunol ; 162(6): 3541-8, 1999 Mar 15.
Article in English | MEDLINE | ID: mdl-10092812

ABSTRACT

Resistance to the mouse pneumonitis (MoPn) strain of Chlamydia trachomatis has been mapped to MHC class II-restricted, IL-12-dependent CD4+ T cells that secrete a type 1 profile of proinflammatory cytokines, which includes IFN-gamma and TNF-alpha. The relative contribution of IFN-gamma is controversial, however, due to variation in results presented by different laboratories. To determine whether C. trachomatis strain differences contributed to this apparent conflict, the relative resistance of IFN-gamma-deficient mice to murine and human strains of C. trachomatis was compared. All human serovars were much more sensitive to the direct inhibitory actions of IFN-gamma than the MoPn strain. Furthermore, genital clearance of human serovar D in the C57BL/6 mouse was mediated by class II-independent mechanisms that probably involved local production of IFN-gamma by cells of the innate immune system. TNF-alpha also contributed indirectly to host resistance against all strains tested. The differential susceptibility of distinct C. trachomatis strains to effector cytokines such as IFN-gamma could not have been predicted by interstrain biologic variation or by the profile of cytokines stimulated during infection. These findings indicate that strain variation should be considered in situations where related isolates of a given parasite produce conflicting data in models of infection and immunity. They also suggest that stimulation of mucosal IFN-gamma activity is a relevant goal for a human chlamydial vaccine.


Subject(s)
Chlamydia Infections/immunology , Chlamydia trachomatis/immunology , Interferon-gamma/pharmacology , Animals , Antibodies, Bacterial/biosynthesis , Chlamydia Infections/microbiology , Chlamydia trachomatis/growth & development , Chlamydia trachomatis/isolation & purification , Cytokines/biosynthesis , Drug Resistance, Microbial , Female , Humans , Immunity, Innate , Mice , Mice, Inbred C57BL , Mice, Inbred Strains , Mice, Knockout , Mucous Membrane/immunology , Mucous Membrane/microbiology , Nitric Oxide/physiology , Pneumonia, Bacterial/immunology , Tumor Necrosis Factor-alpha/physiology , Vaginosis, Bacterial/immunology , Vaginosis, Bacterial/microbiology
6.
J Exp Med ; 188(5): 809-18, 1998 Sep 07.
Article in English | MEDLINE | ID: mdl-9730883

ABSTRACT

Chlamydia trachomatis, an obligate intracellular bacterial pathogen of mucosal surfaces, is a major cause of preventable blindness and sexually transmitted diseases for which vaccines are badly needed. Despite considerable effort, antichlamydial vaccines have proven to be elusive using conventional immunization strategies. We report the use of murine bone marrow-derived dendritic cells (DC) pulsed ex vivo with killed chlamydiae as a novel approach to vaccination against chlamydial infection. Our results show that DC efficiently phagocytose chlamydiae, secrete IL-12 p40, and present chlamydial antigen(s) to infection sensitized CD4(+) T cells. Mice immunized intravenously with chlamydial-pulsed DC produce protective immunity against chlamydial infection of the female genital tract equal to that obtained after infection with live organisms. Immunized mice shed approximately 3 logs fewer infectious chlamydiae and are protected from genital tract inflammatory and obstructive disease. Protective immunity is correlated with a chlamydial-specific Th1-biased response that closely mimics the immune response produced after chlamydial infection. Thus, ex vivo antigen-pulsed DC represent a powerful tool for the study of protective immunity to chlamydial mucosal infection and for the identification of chlamydial protective antigens through reconstitution experiments. Moreover, these findings might impact the design of vaccine strategies against other medically important sexually transmitted diseases for which vaccines are sought but which have proven difficult to develop.


Subject(s)
Adoptive Transfer/methods , Bacterial Vaccines/immunology , Chlamydia Infections/prevention & control , Chlamydia trachomatis/immunology , Dendritic Cells/immunology , Vaginal Diseases/prevention & control , Animals , Antigen Presentation , Antigens, Bacterial/metabolism , CD4-Positive T-Lymphocytes/immunology , CD4-Positive T-Lymphocytes/microbiology , Cells, Cultured , Chlamydia Infections/immunology , Cytokines/metabolism , Dendritic Cells/metabolism , Dendritic Cells/microbiology , Dendritic Cells/transplantation , Female , HeLa Cells , Humans , Mice , Mice, Inbred C57BL , Phagocytosis , Vaccines, Inactivated/immunology , Vaginal Diseases/immunology
7.
J Parasitol ; 80(1): 67-72, 1994 Feb.
Article in English | MEDLINE | ID: mdl-7905921

ABSTRACT

Spleen cells from mice immunized with Cryptosporidium parvum were enriched for T cells by passage over an affinity chromatography column. The proliferative response of these cells was > 2-fold higher than the response of unenriched cells. T-enriched cells were enriched further for either CD4+ cells or CD8+ cells. The proliferative response of CD4-enriched cells was > 4-fold higher than the response by unenriched cells. CD8+ cells were essentially nonresponsive to C. parvum antigen. Culture supernatant fractions from these variously enriched splenocyte populations were assayed for cytokine production. Cultures containing CD4+ cells produced gamma interferon and interleukin-2 following incubation with C. parvum antigen. None of the cultures produced interleukin-4. Production of gamma interferon and interleukin-2, but not interleukin-4, is characteristic of the previously described Th1 helper cell subset. Our data indicate that a subset of murine lymphocytes consistent with the Th1 helper cell phenotype proliferates following in vitro stimulation with C. parvum antigen.


Subject(s)
Antigens, Protozoan/immunology , CD4-Positive T-Lymphocytes/immunology , Cryptosporidium parvum/immunology , Interferon-gamma/biosynthesis , Lymphocyte Activation , Animals , Female , Mice , Mice, Inbred BALB C , Spleen/cytology , Spleen/immunology , T-Lymphocyte Subsets/immunology
8.
Infect Immun ; 61(4): 1460-7, 1993 Apr.
Article in English | MEDLINE | ID: mdl-8454350

ABSTRACT

Lymphocyte blastogenesis assays and immunoblotting were used to investigate and compare murine B-cell responses to preparations of extracellular membrane blebs (BAg) and spirochetes (Ag) of Borrelia burgdorferi. Immunoblotting BAg, Ag, and medium control preparations with serum from naive and infected C57BL/10 mice revealed that BAg and Ag had similar specific reactivity profiles except that major antigens of 83, 60, and 41 kDa were detected in Ag but not in BAg. It was determined that 1 microgram (dry weight) of Ag contained 0.0051 and 0.0063 microgram of outer surface proteins A (OspA) and OspB, respectively, whereas 1 microgram (dry weight) of BAg contained 0.0024 microgram of OspA and 0.0015 microgram of OspB. Both BAg and Ag caused blastogenesis in cultures of spleen cells from both groups of mice, but BAg-stimulated lymphocytes exhibited significantly greater (P < or = 0.05) blastogenesis after 2 or 6 days of culture than did lymphocytes stimulated by Ag or medium control. Flow cytometry and antibody capture enzyme-linked immunosorbent assays identified responding lymphocytes as B cells which secreted polyclonal immunoglobulin M (IgM) but not IgG or IgA. Treatment of BAg and lipopolysaccharide controls with polymyxin B resulted in as much as 20.7 and 54.3% mean decreases in blastogenesis, respectively. Fractionation of BAg or Ag by ultracentrifugation before culture with spleen cells from naive mice indicated that B-cell blastogenesis was probably associated with spirochetal membranes. The results of this study demonstrate that specific humoral responses are directed towards extracellular membrane blebs which lack the 83-, 60-, and 41-kDa antigens of intact spirochetes and that blebs also possess significant nonspecific mitogenic activity for murine B cells. This activity was not due entirely to typical lipopolysaccharide or OspA and OspB lipoproteins.


Subject(s)
B-Lymphocytes/immunology , Borrelia burgdorferi Group/immunology , Lyme Disease/immunology , Animals , Antigens, Bacterial/chemistry , Antigens, Bacterial/immunology , B-Lymphocyte Subsets/cytology , B-Lymphocyte Subsets/immunology , Blotting, Western , Cell Membrane/immunology , Cell Membrane/ultrastructure , Dose-Response Relationship, Immunologic , Female , Lymphocyte Activation , Mice , Mice, Inbred C57BL , Molecular Weight , Polymyxin B/pharmacology , T-Lymphocyte Subsets/cytology , T-Lymphocyte Subsets/immunology
9.
Infect Immun ; 59(3): 990-5, 1991 Mar.
Article in English | MEDLINE | ID: mdl-1997443

ABSTRACT

Cellular and serum antibody responses of calves were monitored for 23 days after oral inoculation of the calves with oocysts of Cryptosporidium parvum. In vitro blastogenic responses of peripheral blood lymphocytes were assessed after stimulation with a C. parvum preparation. Specific lymphocyte blastogenic responses to the parasite were detected 2 days after inoculation. Parasite-specific antibody titers were demonstrable 7 days after inoculation with oocysts and achieved peak levels 9 days after inoculation, coinciding with oocyst shedding at 5 to 10 days after inoculation. Both lymphocyte and antibody responses remained elevated until the termination of the experiment. Immunoblotting the C. parvum preparation with serum from an infected calf revealed six major parasite antigens. Five of these antigens reacted on immunoblots from 7 to 14 days after inoculation with oocysts. A parasite antigen of approximately 11,000 molecular weight demonstrated intense reactivity on immunoblots from 7 to 23 days after inoculation. The 11,000-molecular-weight antigen also reacted on immunoblots with parenterally raised antioocyst and antisporozoite rabbit sera. These results indicate that cell-mediated as well as humoral immune responses are initiated by cryptosporidial infection in calves and that the 11,000-molecular-weight parasite antigen is immunodominant.


Subject(s)
Antibodies, Protozoan/biosynthesis , Cryptosporidiosis/immunology , Lymphocytes/immunology , Animals , Antigens, Protozoan/immunology , Cattle , Electrophoresis, Polyacrylamide Gel , Immunoblotting , Lymphocyte Activation/immunology , Parasite Egg Count
10.
J Parasitol ; 77(1): 170-2, 1991 Feb.
Article in English | MEDLINE | ID: mdl-1992090

ABSTRACT

The ability of murine lymphoid cells or culture supernatant fractions to transfer protection against Cryptosporidium parvum was examined. Spleen or mesenteric lymph node (MLN) cells were taken from adult mice resistant to C. parvum and given either directly or following in vitro culture to infant mice. Neither spleen or MLN cells, nor cells or supernatant fractions from in vitro cultures transferred protection from resistant adult donors to susceptible infant recipients. These results may be due to limitations in the present model. Alternatively, the resistance of infant mice to C. parvum may not be immunologically mediated.


Subject(s)
Cryptosporidiosis/immunology , Immunotherapy, Adoptive , Lymph Nodes/immunology , Spleen/immunology , Animals , Disease Susceptibility , Female , Lymph Nodes/cytology , Male , Mice , Mice, Inbred BALB C , Spleen/cytology
11.
J Parasitol ; 76(3): 450-2, 1990 Jun.
Article in English | MEDLINE | ID: mdl-1693675

ABSTRACT

Spleen and mesenteric lymph node lymphocytes from both Cryptosporidium parvum-exposed and unexposed mice were cultured with antigen (Ag) prepared from C. parvum oocysts. Spleen lymphocytes from oral-, intraperitoneal-, or oral + intraperitoneal-exposed mice did not respond significantly (P greater than 0.05) to Ag stimulation. Spleen lymphocytes from multioral-exposed mice, however, demonstrated significant (P less than or equal to 0.01) Ag-specific blastogenesis. Mesenteric lymph node lymphocytes did not respond to in vitro Ag stimulation regardless of the route of in vivo priming. These results demonstrate an in vitro cell-mediated immune response against C. parvum by lymphocytes in murine spleen.


Subject(s)
Antigens, Protozoan/immunology , Coccidia/immunology , Cryptosporidium/immunology , Lymphocyte Activation , Animals , Cells, Cultured , Cryptosporidiosis/immunology , Epitopes/immunology , Female , Immunity, Cellular , Lymph Nodes/cytology , Male , Mesentery , Mice , Mice, Inbred BALB C , Spleen/cytology
12.
Infect Immun ; 57(3): 999-1001, 1989 Mar.
Article in English | MEDLINE | ID: mdl-2645217

ABSTRACT

P20 is an immunodominant surface antigen of Eimeria bovis sporozoites. As parasites underwent merogony within cultured bovine monocytes and Madin-Darby bovine kidney (MDBK) cells, P20 appeared to be shed gradually by meronts and was absent in type 1 and 2 first-generation merozoites. Meronts of E. bovis appeared to shed P20 into the parasitophorous vacuole of bovine monocytes, whereas MDBK cells evidently released P20 into the culture medium or destroyed its antigenic determinant.


Subject(s)
Antigens, Protozoan/immunology , Antigens, Surface/immunology , Eimeria/immunology , Animals , Eimeria/growth & development , Fluorescent Antibody Technique
13.
J Parasitol ; 75(1): 86-91, 1989 Feb.
Article in English | MEDLINE | ID: mdl-2783968

ABSTRACT

Cellular and humoral responses were investigated following gavage inoculation of 6-wk-old bull calves with 35,000-40,000 oocysts of Eimeria bovis. At 3-4-day intervals for 40 days after inoculation (DAI), blood was taken and assessed for serum IgG against merozoites and sporozoites of E. bovis. Proliferative responses of peripheral blood lymphocytes were measured following stimulation with either concanavalin A (Con A) or a soluble antigen derived from E. bovis oocysts (EbAg). Serum IgG against merozoites and sporozoites reached a peak of activity between 10 and 20 DAI, coinciding with oocyst shedding on days 17 to 24. Serum antibody titers had dropped to base levels by 40 DAI, although anti-merozoite titers remained elevated for the duration of the study (i.e., from days 12 and 20 to day 40). Con A stimulation of lymphocytes was not affected by infection; there was no evidence of suppressed or augmented responsiveness. Lymphocyte responses to EbAg had reached a maximum by day 20 and remained elevated throughout the study. These results indicate (a) that sporozoites and merozoites share antigens recognized by serum IgG, (b) that there is no episode of marked immunosuppression during acute infection, and (c) that cellular immunity is probably more important in resistance against reinfection than humoral immunity.


Subject(s)
Cattle Diseases/immunology , Coccidiosis/veterinary , Acute Disease/immunology , Animals , Antibodies, Protozoan/biosynthesis , Antigens, Protozoan/immunology , Cattle , Coccidiosis/immunology , Concanavalin A/pharmacology , Eimeria/immunology , Immunoglobulin G/biosynthesis , Interleukin-2/biosynthesis , Lymphocyte Activation , Male , T-Lymphocytes/immunology
14.
Infect Immun ; 57(1): 289-90, 1989 Jan.
Article in English | MEDLINE | ID: mdl-2909491

ABSTRACT

Autoradiography identified six 125I-labeled proteins, ranging in molecular weight (Mr) from 20,000 to approximately 110,000, on the plasmalemma of Eimeria bovis sporozoites. Immunoblotting with bovine antiserum generated by intravenous inoculations of sporozoites and with immune serum generated by per os inoculations of oocysts revealed that protein 20 (i.e., 20,000 Mr) was an immunodominant antigen on the surface of E. bovis sporozoites.


Subject(s)
Antigens, Protozoan/isolation & purification , Antigens, Surface/isolation & purification , Eimeria/immunology , Membrane Proteins/isolation & purification , Animals , Antigens, Protozoan/immunology , Antigens, Surface/immunology , Blotting, Western , Cattle , Eimeria/growth & development , Male , Membrane Proteins/immunology , Molecular Weight
15.
Infect Immun ; 56(10): 2538-43, 1988 Oct.
Article in English | MEDLINE | ID: mdl-3047060

ABSTRACT

Five monoclonal antibodies (MAbs) were partially characterized and tested for their ability to inhibit penetration of Madin-Darby bovine kidney (MDBK) cells by sporozoites of Eimeria bovis. By indirect fluorescent-antibody assays, all MAbs reacted with acetone-fixed sporozoites, but only two MAbs, EbS9 (immunoglobulin G1) and EbS11 (immunoglobulin G2a), localized specifically on the plasmalemma of live sporozoites. Two of the five MAbs also reacted with acetone-fixed first-generation merozoites of E. bovis; however, none of the MAbs reacted with live merozoites. Treatment of live sporozoites with EbS9 or EbS11 resulted in 79 and 73% decreases, respectively, in sporozoite penetration of MDBK cells. No significant differences in cell penetration occurred in MDBK cells inoculated with sporozoites that had been treated with the other three MAbs. Both EbS9 and EbS11 reacted in Western blots (immunoblots) of sporozoites with the same 20,000-relative-molecular-weight protein. The antigens against which these neutralizing MAbs react might be useful in immunizing against bovine coccidiosis.


Subject(s)
Antibodies, Monoclonal/immunology , Antibodies, Protozoan/immunology , Antigens, Protozoan/immunology , Antigens, Surface/immunology , Eimeria/immunology , Animals , Cell Line , Cells/parasitology , Dogs , Fluorescent Antibody Technique , Immunoglobulin G/immunology , Mice , Molecular Weight
16.
J Parasitol ; 72(5): 677-83, 1986 Oct.
Article in English | MEDLINE | ID: mdl-3100756

ABSTRACT

Several established cell lines were tested for their ability to support in vitro development of meronts of Sarcocystis cruzi. Sporozoites penetrated bovine monocytes (BM), bovine pulmonary artery endothelial cells (CPA), Madin-Darby bovine kidney cells and mouse macrophages, but developed to meronts in BM and CPA only. Sporozoites developed to large meronts that contained approximately 180-350 merozoites, whereas merozoites formed small meronts with 50-100 merozoites. Mature large meronts were present at 18-86 days after inoculation (DAI) in BM and at 16-72 DAI in CPA. Small meronts were present at 23-115 and 23-91 DAI in BM and CPA. Considerably more merozoites developed in CPA than in BM. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that merozoites harvested at 36 and 48 DAI each had 1 unique protein as well as numerous common proteins.


Subject(s)
Sarcocystis/growth & development , Animals , Cattle , Cell Line , Endothelium/parasitology , Kidney/parasitology , Macrophages/parasitology , Mice , Monocytes/parasitology , Proteins/analysis , Pulmonary Artery , Sarcocystis/analysis , Sarcocystis/cytology
17.
Infect Immun ; 50(2): 566-71, 1985 Nov.
Article in English | MEDLINE | ID: mdl-3876996

ABSTRACT

Sporozoites of Eimeria bovis penetrated and developed normally to first-generation meronts in bovine monocytes (BM) and Madin-Darby bovine kidney (MDBK) cells that had been pretreated with culture medium (CM) or supernatant (NS) from nonstimulated bovine T cells. At 240 h after sporozoite inoculation (ASI), the mean percent development (meronts/[sporozoites + meronts]) in CM- and NS-pretreated BM was 52 and 28%, respectively; values for MDBK cells were 36 and 35%, respectively. Pretreatment of BM and MDBK cells with supernatant (ConAS) from concanavalin A-stimulated bovine T cells had no effect on the ability of sporozoites to penetrate cells; however, at 240 h ASI, only 1% of the sporozoites in ConAS-pretreated BM cultures had developed to meronts. In contrast, ConAS had no adverse effect on the ability of E. bovis sporozoites to develop to first-generation meronts in MDBK cells. At 240 h ASI, E. bovis meronts in ConAS-pretreated BM were abnormal in appearance and retarded in development, whereas sporozoites appeared structurally normal by light microscopy. Pretreatment of BM with ConAS had no effect on the ability of sporozoites of Eimeria papillata (Apicomplexa) to penetrate cells. Sporozoites of E. papillata did not develop to meronts in ConAS-pretreated BM and, in contrast to E. bovis, most sporozoites were destroyed intracellularly.


Subject(s)
Eimeria/growth & development , Lymphocytes/physiology , Lymphokines/pharmacology , Monocytes/physiology , Animals , Cattle , Cell Line , Culture Media , Eimeria/drug effects , Interleukin-2 , Kidney , Species Specificity , T-Lymphocytes/physiology
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