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The expulsion of Echinostoma trivolvis caused by goblet cell hyperplasia in severe combined immunodeficient (SCID) mice
Oral Sessions I Parasitology International 47 (Suppl.) (1998) 133-281
268
49. Immunology-10 (Helminthic Infections)
0-0542
INTESTINAL MASTOCYTOSIS AND GOBLET CELL HYPERI’LASIA IN TWO STRAINS OF MICE INFECTED WITH NEODZPLOSTOMUM
o-o544
IMPAIRED PROTECTION IN CD45EXON6 DEFICIENT MICE AGAINST STRONGYROlDi3 RAl7-1
SEOULENSE &&&g&*.
Chai JY, Kim TK’, Cho WH’, Sea M’, Kook J’, Guk SM’, Lee SH’ ‘Department of Parasitology and Institute of Endemic Diseases, Seoul National University College of Medicine, Seoul 110-799, Korea Mucosal mast cell (MMC) and goblet cell (GC) responses were studied in the small intestine of BALB/c and C3H mice infected with Neodiplosbmum seottlense, and their role in the worm expulsion was studied. During day 3 and day 28 post-infection (PI) with 2CO metacercariae, the worm recovery rate from BALB/c mice was remarkably higher than that from C3H mice. In the duodenum, the main fluke habitat, mastocytosis was pronounced at day 7 PI but quickly diminished thereafter. These MMC kinetics were not different between the two strains of mice. Similar results were observed in the jejunum and ileum, but the extent of mastocytosis was lesser in the ileum. CC hyperplasia was remarkable in the duodenum of BALB/c mice through the course of infection except day 14 PI, whereas it was recognizable only in the jejunum and ileum of C3H mice on day 7 PI. Mucin activation was evidently demonstrated in both strains of mice through the course of infection, but more remarkable in BALB/c than in WH mice. The results strongly suggest that mastocytosis and GC hyperplasia are local immune responses against N. seoulense, however, they play a minor role in the host defense and worm expulsion. o-0543
THE EXPULSION OF ECHINOSTOMA TRIVOLVIS CAUSED BY GOBLET CELL HYPERPLASIA IN SEVERE COMBINED IMMUNODEFICIENT (SCID) MICE
FujinoT.*, *Devent
Ichikawa, H.**, Fukuda, K.***, Fried, B.**** of Biology, Faculty of Science, Yamagata
University, 990 Yamagata, Japan **Department of Medical Zoology. Kanazawa Medical University, 930-02 Ishikawa. Japan, ***Center for Laboratory Ammal Science, National Defence Medical College, 359 Tokorozawa, Japan and ****Department of Biology, Lafayette College, Easton, PA 18042, U.S.A. Mice with severe combined immunodeticiecy (SCID)were each infected with 40 F.&nostoma trivolvis metacercariae on day 0. The test group mice, were given intramuscular injections of dexamethasone (DEX) daily for 2 weeks and necropsied on days 5.8, 12, 15, 20 and 30 p. i. In the controls without the inJection of DEX. worm rejection began about day 8 p. i. and the worms were rejected by day 15 p. i., corresponding to the peak in goblet cell hype@asia, about day 12 p. i. In the test mice, goblet cell hyperplasia was suppressed and the worms were retained until day 15 p. i., and then rejected after the last treatment with DEX. The number of mucosal mast cells in the controls, peaked about day 15 p, i., was suppressed with DEX. The eosmophil number m the controls increased about on day 15 p. 1. and then decreased. The eosmophil number in the test nuce was significantly suppressed compared to that of the controls dunng the period of the experiment. Enzyme-linked immunosorbent assay @LISA) showed no marked rise in titres of IgM, IgA and IgG throughout the experiment in both groups. These results mdicate that DEXtreatment delayed the rejection of 5. trivolvis from SCID mice m assoclatlon
with the suppression
of goblet cell hyperplasla.
Hamano S*. Kishihara K**, Nomoto K**, Tada I*
* Department of Parasitology, Faculty of Medicine ** Department of Immunology, Medical Institute of Bioregulation Kyushu University, Fukuoka, Japan CD45 is a membrane tyrosine phosphatase and its necessity has been shown in various cells. It has also been shown that CD45 is necessary for IgE dependent degranulation of mast cells. To examine whether CD45 is essential for development and/or activation of mucosal mast cell (MMC), we studied the host protective immunity against intestinal nematoda,Strongyfoides ratti (Sr), in CD45exod deficient mice (CD45-/-, C57BU6 background), since the expulsion of Sr is dependent on MMC. CD45 -/-and +/+ were subcutaneously infected with 2000 third stage larvae (L3) of Sr. The number of MMC and eggs per gram feces (EFG) were monitored. In both groups, the number of MMC before infection were similar and the peaks of EPG were observed on day 6 after infection. No egg was detected in CD45 +/+ on day 14, while significant eggs were still detected in CD45/-. It took 40 days after infection to clear the eggs in CD45-/-. As this persistent infection may be due to impaired T cell function in CD45-/-, we transferred normal spleen cells (8 X lol/mouse) to CD45-/-. 6 days after transfer, CD45/were infected with 2OOOL3 of Sr. On 2 1 days after infection, eggs were still detected from the reconstituted mice. Nextly we injected IL-3 (2OCXlU/day) to CD45-/- during day 6 to 10. On day 2 1.eggs were detected. MMC numbers of the day were as follows; CD45-/6.6/1OVCU, CD45-/- +IL-3 24/1OVCU and control 27/1OVCU. After induction of MMC with &3,CD45-/couldn’t exclude Sr. CD45-/have unknown defects in protection mechanism against Sr infection. O-0545
INDUCTION OF PROTECTIVE IMMUNITY AGAINST ONCHOCERCA VOLVLJLUS LARVAE BY IMMUNIZATION WITH RECOMBINANT PROTEINS
Lustigrnan S*, Joseph GT*, Huima T’, Abraham D” lLindsiey F. Kimball Reseuch Institute, New York Blood Center, New York, NY and **Thomas Jefferson University, Philadelphia, PA Antibody and cell-dependent immune mechanisms may contribute to resistance to larval 0. volv&.s in humans and in animal models. In our effort to identify and clone larval proteins that can induce protective immunity to 0. volvu/us larvae, we immunosereened 0. volvulur cDNA expression libraries with sera from chimpanzees that were immunized with X-irradiated larvae, and with a pool of sem from putatively protected individuals (PI). Three of the recombinant antigens (OV7, 0v64 and OvB8) that were selected for testing as vaccine candidates were found by immunolocalization analysis to he expressed in L3 and during the development of L3 to L4. Analysis of the antibody responses (IgG and IgE) to the recombinant proteins in the PI in comparison to the infected individuals have shown that the recombinant antigens were recognized equally by both groups. For vaccination trials, mice we= immunized in the presence of alum and the efficacy of immunization was eyaluated by recovery of 0. volvultls larvae from diffusion chambers implanted subcutaneously. Reduction of challenge worm survival compared with controls of approximately 45% was induced by immunization with GST-OV7, 0V7, GST-0~64, and with GSTOvB8. Although the mechanism of protective immunity induced in mice hy these antigens has yet to be determined, we have observed that human monospecific anti-OV7 antibodies completely inhibited the molting of L3 to L4 in vitro in the presence of neutrophils. Studies are currently in progress to optimize immunization with these recombinant antigens, to determine the immune mechanisms operating in mice and to compare these findings to those obtained for humans in viva and in vitro.
268
49. Immunology-10 (Helminthic Infections)
0-0542
INTESTINAL MASTOCYTOSIS AND GOBLET CELL HYPERI’LASIA IN TWO STRAINS OF MICE INFECTED WITH NEODZPLOSTOMUM
o-o544
IMPAIRED PROTECTION IN CD45EXON6 DEFICIENT MICE AGAINST STRONGYROlDi3 RAl7-1
SEOULENSE &&&g&*.
Chai JY, Kim TK’, Cho WH’, Sea M’, Kook J’, Guk SM’, Lee SH’ ‘Department of Parasitology and Institute of Endemic Diseases, Seoul National University College of Medicine, Seoul 110-799, Korea Mucosal mast cell (MMC) and goblet cell (GC) responses were studied in the small intestine of BALB/c and C3H mice infected with Neodiplosbmum seottlense, and their role in the worm expulsion was studied. During day 3 and day 28 post-infection (PI) with 2CO metacercariae, the worm recovery rate from BALB/c mice was remarkably higher than that from C3H mice. In the duodenum, the main fluke habitat, mastocytosis was pronounced at day 7 PI but quickly diminished thereafter. These MMC kinetics were not different between the two strains of mice. Similar results were observed in the jejunum and ileum, but the extent of mastocytosis was lesser in the ileum. CC hyperplasia was remarkable in the duodenum of BALB/c mice through the course of infection except day 14 PI, whereas it was recognizable only in the jejunum and ileum of C3H mice on day 7 PI. Mucin activation was evidently demonstrated in both strains of mice through the course of infection, but more remarkable in BALB/c than in WH mice. The results strongly suggest that mastocytosis and GC hyperplasia are local immune responses against N. seoulense, however, they play a minor role in the host defense and worm expulsion. o-0543
THE EXPULSION OF ECHINOSTOMA TRIVOLVIS CAUSED BY GOBLET CELL HYPERPLASIA IN SEVERE COMBINED IMMUNODEFICIENT (SCID) MICE
FujinoT.*, *Devent
Ichikawa, H.**, Fukuda, K.***, Fried, B.**** of Biology, Faculty of Science, Yamagata
University, 990 Yamagata, Japan **Department of Medical Zoology. Kanazawa Medical University, 930-02 Ishikawa. Japan, ***Center for Laboratory Ammal Science, National Defence Medical College, 359 Tokorozawa, Japan and ****Department of Biology, Lafayette College, Easton, PA 18042, U.S.A. Mice with severe combined immunodeticiecy (SCID)were each infected with 40 F.&nostoma trivolvis metacercariae on day 0. The test group mice, were given intramuscular injections of dexamethasone (DEX) daily for 2 weeks and necropsied on days 5.8, 12, 15, 20 and 30 p. i. In the controls without the inJection of DEX. worm rejection began about day 8 p. i. and the worms were rejected by day 15 p. i., corresponding to the peak in goblet cell hype@asia, about day 12 p. i. In the test mice, goblet cell hyperplasia was suppressed and the worms were retained until day 15 p. i., and then rejected after the last treatment with DEX. The number of mucosal mast cells in the controls, peaked about day 15 p, i., was suppressed with DEX. The eosmophil number m the controls increased about on day 15 p. 1. and then decreased. The eosmophil number in the test nuce was significantly suppressed compared to that of the controls dunng the period of the experiment. Enzyme-linked immunosorbent assay @LISA) showed no marked rise in titres of IgM, IgA and IgG throughout the experiment in both groups. These results mdicate that DEXtreatment delayed the rejection of 5. trivolvis from SCID mice m assoclatlon
with the suppression
of goblet cell hyperplasla.
Hamano S*. Kishihara K**, Nomoto K**, Tada I*
* Department of Parasitology, Faculty of Medicine ** Department of Immunology, Medical Institute of Bioregulation Kyushu University, Fukuoka, Japan CD45 is a membrane tyrosine phosphatase and its necessity has been shown in various cells. It has also been shown that CD45 is necessary for IgE dependent degranulation of mast cells. To examine whether CD45 is essential for development and/or activation of mucosal mast cell (MMC), we studied the host protective immunity against intestinal nematoda,Strongyfoides ratti (Sr), in CD45exod deficient mice (CD45-/-, C57BU6 background), since the expulsion of Sr is dependent on MMC. CD45 -/-and +/+ were subcutaneously infected with 2000 third stage larvae (L3) of Sr. The number of MMC and eggs per gram feces (EFG) were monitored. In both groups, the number of MMC before infection were similar and the peaks of EPG were observed on day 6 after infection. No egg was detected in CD45 +/+ on day 14, while significant eggs were still detected in CD45/-. It took 40 days after infection to clear the eggs in CD45-/-. As this persistent infection may be due to impaired T cell function in CD45-/-, we transferred normal spleen cells (8 X lol/mouse) to CD45-/-. 6 days after transfer, CD45/were infected with 2OOOL3 of Sr. On 2 1 days after infection, eggs were still detected from the reconstituted mice. Nextly we injected IL-3 (2OCXlU/day) to CD45-/- during day 6 to 10. On day 2 1.eggs were detected. MMC numbers of the day were as follows; CD45-/6.6/1OVCU, CD45-/- +IL-3 24/1OVCU and control 27/1OVCU. After induction of MMC with &3,CD45-/couldn’t exclude Sr. CD45-/have unknown defects in protection mechanism against Sr infection. O-0545
INDUCTION OF PROTECTIVE IMMUNITY AGAINST ONCHOCERCA VOLVLJLUS LARVAE BY IMMUNIZATION WITH RECOMBINANT PROTEINS
Lustigrnan S*, Joseph GT*, Huima T’, Abraham D” lLindsiey F. Kimball Reseuch Institute, New York Blood Center, New York, NY and **Thomas Jefferson University, Philadelphia, PA Antibody and cell-dependent immune mechanisms may contribute to resistance to larval 0. volv&.s in humans and in animal models. In our effort to identify and clone larval proteins that can induce protective immunity to 0. volvu/us larvae, we immunosereened 0. volvulur cDNA expression libraries with sera from chimpanzees that were immunized with X-irradiated larvae, and with a pool of sem from putatively protected individuals (PI). Three of the recombinant antigens (OV7, 0v64 and OvB8) that were selected for testing as vaccine candidates were found by immunolocalization analysis to he expressed in L3 and during the development of L3 to L4. Analysis of the antibody responses (IgG and IgE) to the recombinant proteins in the PI in comparison to the infected individuals have shown that the recombinant antigens were recognized equally by both groups. For vaccination trials, mice we= immunized in the presence of alum and the efficacy of immunization was eyaluated by recovery of 0. volvultls larvae from diffusion chambers implanted subcutaneously. Reduction of challenge worm survival compared with controls of approximately 45% was induced by immunization with GST-OV7, 0V7, GST-0~64, and with GSTOvB8. Although the mechanism of protective immunity induced in mice hy these antigens has yet to be determined, we have observed that human monospecific anti-OV7 antibodies completely inhibited the molting of L3 to L4 in vitro in the presence of neutrophils. Studies are currently in progress to optimize immunization with these recombinant antigens, to determine the immune mechanisms operating in mice and to compare these findings to those obtained for humans in viva and in vitro.