Pub Date : 2001-11-01DOI: 10.1128/CDLI.8.6.1056-1059.2001
E. Ishii, K. Yokota, T. Sugiyama, Y. Fujinaga, K. Ayada, I. Hokari, S. Hayashi, Y. Hirai, M. Asaka, K. Oguma
ABSTRACT Gastric mucosa-associated lymphoid tissue (MALT) lymphoma is related to Helicobacter pylori infection. Specifically, it has been pointed out that pathogenesis of MALT lymphoma involves the 60-kDa heat shock protein (hsp60). To investigate humoral immune responses to the H. pylori hsp60 in patients with gastroduodenal diseases and patients with MALT lymphoma, the hsp60 ofH. pylori was expressed with a glutathioneS-transferase fusion protein and was purified (recombinant hsp60). Sera were obtained from H. pylori-positive patients with gastroduodenal diseases (MALT lymphoma, n = 13; gastric ulcer, n = 20; duodenal ulcer, n = 20; gastritis,n = 20) and from H. pylori-negative healthy volunteers (n = 9). Sera from patients with MALT lymphoma were also obtained at two times: before and after eradication therapy. Antibodies to hsp60 and H. pylori were assessed by enzyme-linked immunosorbent assay. The levels of immunoglobulin G (IgG) antibodies to the hsp60 of H. pylori-positive patients with gastroduodenal diseases were significantly elevated compared to those in the controls. The levels of IgG1 antibodies to hsp60 were elevated and correlated with the levels of anti-H. pylori antibodies in patients with MALT lymphoma. Humarol immunity against hsp60 may be important and relevant to gastroduodenal diseases induced by H. pylori infection.
{"title":"Immunoglobulin G1 Antibody Response toHelicobacter pylori Heat Shock Protein 60 Is Closely Associated with Low-Grade Gastric Mucosa-Associated Lymphoid Tissue Lymphoma","authors":"E. Ishii, K. Yokota, T. Sugiyama, Y. Fujinaga, K. Ayada, I. Hokari, S. Hayashi, Y. Hirai, M. Asaka, K. Oguma","doi":"10.1128/CDLI.8.6.1056-1059.2001","DOIUrl":"https://doi.org/10.1128/CDLI.8.6.1056-1059.2001","url":null,"abstract":"ABSTRACT Gastric mucosa-associated lymphoid tissue (MALT) lymphoma is related to Helicobacter pylori infection. Specifically, it has been pointed out that pathogenesis of MALT lymphoma involves the 60-kDa heat shock protein (hsp60). To investigate humoral immune responses to the H. pylori hsp60 in patients with gastroduodenal diseases and patients with MALT lymphoma, the hsp60 ofH. pylori was expressed with a glutathioneS-transferase fusion protein and was purified (recombinant hsp60). Sera were obtained from H. pylori-positive patients with gastroduodenal diseases (MALT lymphoma, n = 13; gastric ulcer, n = 20; duodenal ulcer, n = 20; gastritis,n = 20) and from H. pylori-negative healthy volunteers (n = 9). Sera from patients with MALT lymphoma were also obtained at two times: before and after eradication therapy. Antibodies to hsp60 and H. pylori were assessed by enzyme-linked immunosorbent assay. The levels of immunoglobulin G (IgG) antibodies to the hsp60 of H. pylori-positive patients with gastroduodenal diseases were significantly elevated compared to those in the controls. The levels of IgG1 antibodies to hsp60 were elevated and correlated with the levels of anti-H. pylori antibodies in patients with MALT lymphoma. Humarol immunity against hsp60 may be important and relevant to gastroduodenal diseases induced by H. pylori infection.","PeriodicalId":10395,"journal":{"name":"Clinical Diagnostic Laboratory Immunology","volume":"2 1","pages":"1056 - 1059"},"PeriodicalIF":0.0,"publicationDate":"2001-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73254211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-11-01DOI: 10.1128/CDLI.8.6.1240-1247.2001
K. Marr, Michael Koudadoust, M. Black, S. Arunmozhi Balajee
ABSTRACT Detailed investigations of macrophage phagocytosis and killing ofAspergillus fumigatus conidia have been limited by technical difficulties in quantifying fungal uptake and viability. In order to study early events in cell pathogen ingestion and killing, we developed a new flow cytometry assay that utilizes the fungus-specific viability dye FUN-1. Metabolically active A. fumigatusconidia accumulate orange fluorescence in vacuoles, while dormant or dead conidia stain green. After incubation within THP-1 cells, recovered conidia are costained with propidium iodide (PI) to discriminate between dormant and dead cells. Flow cytometric measurements of FUN-1 metabolism and PI uptake provide indicators of conidial viability, dormancy, and death. Conidial phagocytosis and killing are also assessed by measurement of green and orange FUN-1 fluorescence within the THP-1 cell population. Compared to previously described methods, this assay has less error introduced by membrane permeability changes and serial dilution of filamentous fungal forms. Results suggest that the THP-1 cells kill conidia rapidly (within 6 h) after exposure. Conidia that are preexposed to human serum are ingested and killed more quickly than are nonopsonized conidia.
{"title":"Early Events in Macrophage Killing ofAspergillus fumigatus Conidia: New Flow Cytometric Viability Assay","authors":"K. Marr, Michael Koudadoust, M. Black, S. Arunmozhi Balajee","doi":"10.1128/CDLI.8.6.1240-1247.2001","DOIUrl":"https://doi.org/10.1128/CDLI.8.6.1240-1247.2001","url":null,"abstract":"ABSTRACT Detailed investigations of macrophage phagocytosis and killing ofAspergillus fumigatus conidia have been limited by technical difficulties in quantifying fungal uptake and viability. In order to study early events in cell pathogen ingestion and killing, we developed a new flow cytometry assay that utilizes the fungus-specific viability dye FUN-1. Metabolically active A. fumigatusconidia accumulate orange fluorescence in vacuoles, while dormant or dead conidia stain green. After incubation within THP-1 cells, recovered conidia are costained with propidium iodide (PI) to discriminate between dormant and dead cells. Flow cytometric measurements of FUN-1 metabolism and PI uptake provide indicators of conidial viability, dormancy, and death. Conidial phagocytosis and killing are also assessed by measurement of green and orange FUN-1 fluorescence within the THP-1 cell population. Compared to previously described methods, this assay has less error introduced by membrane permeability changes and serial dilution of filamentous fungal forms. Results suggest that the THP-1 cells kill conidia rapidly (within 6 h) after exposure. Conidia that are preexposed to human serum are ingested and killed more quickly than are nonopsonized conidia.","PeriodicalId":10395,"journal":{"name":"Clinical Diagnostic Laboratory Immunology","volume":"7 1","pages":"1240 - 1247"},"PeriodicalIF":0.0,"publicationDate":"2001-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88755478","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-11-01DOI: 10.1128/CDLI.8.6.1234-1239.2001
M. Yamaguchi, Motoi Matsuura, Kiyoshi Kobayashi, H. Sasaki, T. Yajima, T. Kuwata
ABSTRACT BALB/c mice were intravenously injected with lipopolysaccharide (LPS) (0.05 μg/g of body weight) 7 days after being primed with zymosan. Recombinant human lactoferrin (250 μg/g of body weight), intravenously administered 1 day before the injection of LPS, significantly lessened the severity of hepatitis, as assessed by levels of serum alanine transaminase compared to those seen when casein was administered. The transient rise of serum tumor necrosis factor alpha (TNF-α) after LPS treatment was also significantly lowered by the intravenous administration of lactoferrin, suggesting that the effect of lactoferrin was due to the suppression of TNF-α production. The following results indicate that the sites of action of lactoferrin for the suppression of the development of this type of hepatitis are Kupffer cells. Gadolinium chloride, a substance known to eliminate Kupffer cells, administered 1 day before LPS, inhibited the transient rise of TNF-α and protected against the development of hepatitis. Kupffer cells isolated from mice intraperitoneally injected with recombinant human lactoferrin became refractory to LPS. The specific interaction of recombinant human lactoferrin with the Kupffer cells was shown by a binding assay, which revealed two types of binding sites on mouse Kupffer cells. Of the two dissociation constants determined in this way, the lower dissociation constant, 0.47 × 10−6 M, was within the range of the 50% effective doses for the suppression of TNF-α production. These results suggest that recombinant human lactoferrin administered to mice suppresses the production of TNF-α by Kupffer cells by directly associating with the binding sites on these cells.
{"title":"Lactoferrin Protects against Development of Hepatitis Caused by Sensitization of Kupffer Cells by Lipopolysaccharide","authors":"M. Yamaguchi, Motoi Matsuura, Kiyoshi Kobayashi, H. Sasaki, T. Yajima, T. Kuwata","doi":"10.1128/CDLI.8.6.1234-1239.2001","DOIUrl":"https://doi.org/10.1128/CDLI.8.6.1234-1239.2001","url":null,"abstract":"ABSTRACT BALB/c mice were intravenously injected with lipopolysaccharide (LPS) (0.05 μg/g of body weight) 7 days after being primed with zymosan. Recombinant human lactoferrin (250 μg/g of body weight), intravenously administered 1 day before the injection of LPS, significantly lessened the severity of hepatitis, as assessed by levels of serum alanine transaminase compared to those seen when casein was administered. The transient rise of serum tumor necrosis factor alpha (TNF-α) after LPS treatment was also significantly lowered by the intravenous administration of lactoferrin, suggesting that the effect of lactoferrin was due to the suppression of TNF-α production. The following results indicate that the sites of action of lactoferrin for the suppression of the development of this type of hepatitis are Kupffer cells. Gadolinium chloride, a substance known to eliminate Kupffer cells, administered 1 day before LPS, inhibited the transient rise of TNF-α and protected against the development of hepatitis. Kupffer cells isolated from mice intraperitoneally injected with recombinant human lactoferrin became refractory to LPS. The specific interaction of recombinant human lactoferrin with the Kupffer cells was shown by a binding assay, which revealed two types of binding sites on mouse Kupffer cells. Of the two dissociation constants determined in this way, the lower dissociation constant, 0.47 × 10−6 M, was within the range of the 50% effective doses for the suppression of TNF-α production. These results suggest that recombinant human lactoferrin administered to mice suppresses the production of TNF-α by Kupffer cells by directly associating with the binding sites on these cells.","PeriodicalId":10395,"journal":{"name":"Clinical Diagnostic Laboratory Immunology","volume":"2334 1","pages":"1234 - 1239"},"PeriodicalIF":0.0,"publicationDate":"2001-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86569414","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-11-01DOI: 10.1128/CDLI.8.6.1277-1278.2001
C. F. von Reyn, Paige L. Williams, H. Lederman, J. McCutchan, S. Koletar, R. Murphy, S. Cohn, Thomas Evans, A. Heald, Dodi Colquhoun, E. Bassily, J. Currier
ABSTRACT Skin tests and lymphocyte proliferation assays (LPA) were performed with Mycobacterium avium sensitin on patients with AIDS. Among 139 subjects, 13% had positive skin test results and 32% had positive LPA results. The LPA may be a more sensitive indicator of prior M. avium infection in this population.
{"title":"Skin Test Reactivity and Cellular Immune Responses toMycobacterium avium Sensitin in AIDS Patients at Risk for Disseminated M. avium Infection","authors":"C. F. von Reyn, Paige L. Williams, H. Lederman, J. McCutchan, S. Koletar, R. Murphy, S. Cohn, Thomas Evans, A. Heald, Dodi Colquhoun, E. Bassily, J. Currier","doi":"10.1128/CDLI.8.6.1277-1278.2001","DOIUrl":"https://doi.org/10.1128/CDLI.8.6.1277-1278.2001","url":null,"abstract":"ABSTRACT Skin tests and lymphocyte proliferation assays (LPA) were performed with Mycobacterium avium sensitin on patients with AIDS. Among 139 subjects, 13% had positive skin test results and 32% had positive LPA results. The LPA may be a more sensitive indicator of prior M. avium infection in this population.","PeriodicalId":10395,"journal":{"name":"Clinical Diagnostic Laboratory Immunology","volume":"93 1","pages":"1277 - 1278"},"PeriodicalIF":0.0,"publicationDate":"2001-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85289817","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-11-01DOI: 10.1128/CDLI.8.6.1110-1114.2001
S. Moyo, J. Maeland, J. Mudzori
ABSTRACT Group B streptococci (GBS) express strain-variable and surface-localized proteins, which are important serotype markers and targets of protective antibodies. These include the cαand R4 proteins, one or the other of which is expressed by approximately 75% of clinical GBS isolates. These proteins have been considered vaccine candidates. In this study, the cα and R4 proteins were extracted by trypsin digestion of GBS and purified by sequential precipitation with trichloroacetic acid and ammonium sulfate followed by gel filtration chromatography. The proteins were used as antigens in an indirect enzyme-linked immunosorbent assay (ELISA) to measure the levels of cα- and R4-reactive antibodies in sera from pregnant women from Norway (n = 100) and from Zimbabwe (n = 124). Antibody levels in the Norwegian group of women were significantly higher than in the Zimbabwean group, and a higher proportion of the Norwegian women contained appreciable levels of antibodies against both proteins. The antibodies traversed the placental barrier. With individual sera, a significant correlation between the anti-cα and anti-R4 antibody levels was observed and each of the two protein antigens effectively competed for human serum antibodies both against itself and against the other antigen. Inhibition ELISA results demonstrated specificity for each of the proteins of immune antibodies raised in rabbits. These results demonstrate that (i) the majority of women of childbearing age have antibodies against cα and R4, (ii) the levels of these antibodies differ among pregnant women in different parts of the world, and (iii) the normal human serum antibodies may target a common cα and R4 protein site, whereas immune antibodies targeted a different site(s) specific for each protein.
{"title":"Antibodies against Streptococcus agalactiae Proteins cα and R4 in Sera from Pregnant Women from Norway and Zimbabwe","authors":"S. Moyo, J. Maeland, J. Mudzori","doi":"10.1128/CDLI.8.6.1110-1114.2001","DOIUrl":"https://doi.org/10.1128/CDLI.8.6.1110-1114.2001","url":null,"abstract":"ABSTRACT Group B streptococci (GBS) express strain-variable and surface-localized proteins, which are important serotype markers and targets of protective antibodies. These include the cαand R4 proteins, one or the other of which is expressed by approximately 75% of clinical GBS isolates. These proteins have been considered vaccine candidates. In this study, the cα and R4 proteins were extracted by trypsin digestion of GBS and purified by sequential precipitation with trichloroacetic acid and ammonium sulfate followed by gel filtration chromatography. The proteins were used as antigens in an indirect enzyme-linked immunosorbent assay (ELISA) to measure the levels of cα- and R4-reactive antibodies in sera from pregnant women from Norway (n = 100) and from Zimbabwe (n = 124). Antibody levels in the Norwegian group of women were significantly higher than in the Zimbabwean group, and a higher proportion of the Norwegian women contained appreciable levels of antibodies against both proteins. The antibodies traversed the placental barrier. With individual sera, a significant correlation between the anti-cα and anti-R4 antibody levels was observed and each of the two protein antigens effectively competed for human serum antibodies both against itself and against the other antigen. Inhibition ELISA results demonstrated specificity for each of the proteins of immune antibodies raised in rabbits. These results demonstrate that (i) the majority of women of childbearing age have antibodies against cα and R4, (ii) the levels of these antibodies differ among pregnant women in different parts of the world, and (iii) the normal human serum antibodies may target a common cα and R4 protein site, whereas immune antibodies targeted a different site(s) specific for each protein.","PeriodicalId":10395,"journal":{"name":"Clinical Diagnostic Laboratory Immunology","volume":"2018 1","pages":"1110 - 1114"},"PeriodicalIF":0.0,"publicationDate":"2001-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73576649","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-11-01DOI: 10.1128/CDLI.8.6.1136-1139.2001
Y. Shimazaki, M. Mitoma, T. Oho, Y. Nakano, Y. Yamashita, K. Okano, Yutaka Nakano, M. Fukuyama, N. Fujihara, Y. Nada, T. Koga
ABSTRACT Cell surface protein antigen (PAc) and water-insoluble glucan-synthesizing enzyme (GTF-I) produced by cariogenicStreptococcus mutans are two major factors implicated in the colonization of the human oral cavity by this bacterium. We examined the effect of bovine milk, produced after immunization with a fusion protein of functional domains of these proteins, on the recolonization of S. mutans. To prepare immune milk, a pregnant Holstein cow was immunized with the fusion protein PAcA-GB, a fusion of the saliva-binding alanine-rich region (PAcA) of PAc and the glucan-binding (GB) domain of GTF-I. After eight adult subjects received cetylpyridinium chloride (CPC) treatment, one subgroup (n = 4) rinsed their mouths with immune milk and a control group (n = 4) rinsed with nonimmune milk. S. mutans levels in saliva and dental plaque decreased after CPC treatment in both groups. Mouth rinsing with immune milk significantly inhibited recolonization of S. mutans in saliva and plaque. On the other hand, the numbers of S. mutans cells in saliva and plaque in the control group increased immediately after the CPC treatment and surpassed the baseline level 42 and 28 days, respectively, after the CPC treatment. The ratios ofS. mutans to total streptococci in saliva and plaque in the group that received immune milk were lower than those in the control group. These results suggest that milk produced from immunized cow may be useful for controlling S. mutans in the human oral cavity.
{"title":"Passive Immunization with Milk Produced from an Immunized Cow Prevents Oral Recolonization by Streptococcus mutans","authors":"Y. Shimazaki, M. Mitoma, T. Oho, Y. Nakano, Y. Yamashita, K. Okano, Yutaka Nakano, M. Fukuyama, N. Fujihara, Y. Nada, T. Koga","doi":"10.1128/CDLI.8.6.1136-1139.2001","DOIUrl":"https://doi.org/10.1128/CDLI.8.6.1136-1139.2001","url":null,"abstract":"ABSTRACT Cell surface protein antigen (PAc) and water-insoluble glucan-synthesizing enzyme (GTF-I) produced by cariogenicStreptococcus mutans are two major factors implicated in the colonization of the human oral cavity by this bacterium. We examined the effect of bovine milk, produced after immunization with a fusion protein of functional domains of these proteins, on the recolonization of S. mutans. To prepare immune milk, a pregnant Holstein cow was immunized with the fusion protein PAcA-GB, a fusion of the saliva-binding alanine-rich region (PAcA) of PAc and the glucan-binding (GB) domain of GTF-I. After eight adult subjects received cetylpyridinium chloride (CPC) treatment, one subgroup (n = 4) rinsed their mouths with immune milk and a control group (n = 4) rinsed with nonimmune milk. S. mutans levels in saliva and dental plaque decreased after CPC treatment in both groups. Mouth rinsing with immune milk significantly inhibited recolonization of S. mutans in saliva and plaque. On the other hand, the numbers of S. mutans cells in saliva and plaque in the control group increased immediately after the CPC treatment and surpassed the baseline level 42 and 28 days, respectively, after the CPC treatment. The ratios ofS. mutans to total streptococci in saliva and plaque in the group that received immune milk were lower than those in the control group. These results suggest that milk produced from immunized cow may be useful for controlling S. mutans in the human oral cavity.","PeriodicalId":10395,"journal":{"name":"Clinical Diagnostic Laboratory Immunology","volume":"76 1","pages":"1136 - 1139"},"PeriodicalIF":0.0,"publicationDate":"2001-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81060120","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-11-01DOI: 10.1128/CDLI.8.6.1131-1135.2001
Simona Neri, E. Mariani, A. Meneghetti, L. Cattini, A. Facchini
ABSTRACT Cytotoxicity assays provide an in vitro evaluation of the lytic activity of NK and T cells against tumors or transformed cells. However, none of these methods allow the recovery of cells or supernatants after the assay. We standardized a microcytotoxicity test using calcein-acetoxymethyl (calcein-AM) dye that requires very small quantities of cells while maintaining the same sensitivity as the traditional 51Cr assay. The assay is applicable to resting as well as activated human effector cells and uses different targets such as human cell lines that are adherent or growing in suspension and resistant or sensitive. The most important feature of the method is the possibility of recovering cells and supernatants for additional analyses such as phenotyping and evaluation of soluble factors.
{"title":"Calcein-Acetyoxymethyl Cytotoxicity Assay: Standardization of a Method Allowing Additional Analyses on Recovered Effector Cells and Supernatants","authors":"Simona Neri, E. Mariani, A. Meneghetti, L. Cattini, A. Facchini","doi":"10.1128/CDLI.8.6.1131-1135.2001","DOIUrl":"https://doi.org/10.1128/CDLI.8.6.1131-1135.2001","url":null,"abstract":"ABSTRACT Cytotoxicity assays provide an in vitro evaluation of the lytic activity of NK and T cells against tumors or transformed cells. However, none of these methods allow the recovery of cells or supernatants after the assay. We standardized a microcytotoxicity test using calcein-acetoxymethyl (calcein-AM) dye that requires very small quantities of cells while maintaining the same sensitivity as the traditional 51Cr assay. The assay is applicable to resting as well as activated human effector cells and uses different targets such as human cell lines that are adherent or growing in suspension and resistant or sensitive. The most important feature of the method is the possibility of recovering cells and supernatants for additional analyses such as phenotyping and evaluation of soluble factors.","PeriodicalId":10395,"journal":{"name":"Clinical Diagnostic Laboratory Immunology","volume":"30 1","pages":"1131 - 1135"},"PeriodicalIF":0.0,"publicationDate":"2001-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87599721","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-11-01DOI: 10.1128/CDLI.8.6.1104-1109.2001
M. O'gorman, B. Duchateau, M. Paniagua, J. Hunt, Nicolas Bensen, R. Yogev
ABSTRACT The CD40 ligand (CD154), expressed primarily on activated CD4-positive T cells, is a costimulatory molecule involved in B-cell proliferation, germinal center formation, and immunoglobulin class switching. Since B-cell abnormalities including hypergammaglobulinemia and abnormal antibody-specific immune responses are prominent and occur early during the course of pediatric human immunodeficiency virus (HIV) infection, we measured the baseline levels and the induced levels of expression of CD154 on CD3+ CD8− (T helper cells) in HIV-infected children and uninfected children born to HIV-positive mothers. The percentage of CD154+ T helper cells activated in vitro and the level of CD154 expressed per T helper cell (mean fluorescent channel [MFC]) were significantly lower in the HIV-infected children than in the uninfected control group (77% ± 3% versus 89% ± 1%, respectively [P < 0.002], and 261 ± 174 versus 415 ± 130 MFC, respectively [P < 0.03]). The levels of CD154 expressed on resting T helper cells in the HIV-infected group were not significantly different from the levels observed in the control group. In the HIV-infected children, the level of CD154 on activated T helper cells correlated with the level of immunodeficiency, as assessed by the CD4 T-cell levels (correlation coefficient [r] = 0.707,P = 0.003), but did not correlate with the viral load or with any of the serum immunoglobulin concentrations measured in this group of HIV-infected children. The baseline level of CD154 expressed on T helper cells did, however, correlate with the concentration of immunoglobulin A in serum. We conclude that HIV-infected children have impaired regulation of CD154 expression which may contribute to the immune dysregulation commonly observed.
{"title":"Abnormal CD40 Ligand (CD154) Expression in Human Immunodeficiency Virus-Infected Children","authors":"M. O'gorman, B. Duchateau, M. Paniagua, J. Hunt, Nicolas Bensen, R. Yogev","doi":"10.1128/CDLI.8.6.1104-1109.2001","DOIUrl":"https://doi.org/10.1128/CDLI.8.6.1104-1109.2001","url":null,"abstract":"ABSTRACT The CD40 ligand (CD154), expressed primarily on activated CD4-positive T cells, is a costimulatory molecule involved in B-cell proliferation, germinal center formation, and immunoglobulin class switching. Since B-cell abnormalities including hypergammaglobulinemia and abnormal antibody-specific immune responses are prominent and occur early during the course of pediatric human immunodeficiency virus (HIV) infection, we measured the baseline levels and the induced levels of expression of CD154 on CD3+ CD8− (T helper cells) in HIV-infected children and uninfected children born to HIV-positive mothers. The percentage of CD154+ T helper cells activated in vitro and the level of CD154 expressed per T helper cell (mean fluorescent channel [MFC]) were significantly lower in the HIV-infected children than in the uninfected control group (77% ± 3% versus 89% ± 1%, respectively [P < 0.002], and 261 ± 174 versus 415 ± 130 MFC, respectively [P < 0.03]). The levels of CD154 expressed on resting T helper cells in the HIV-infected group were not significantly different from the levels observed in the control group. In the HIV-infected children, the level of CD154 on activated T helper cells correlated with the level of immunodeficiency, as assessed by the CD4 T-cell levels (correlation coefficient [r] = 0.707,P = 0.003), but did not correlate with the viral load or with any of the serum immunoglobulin concentrations measured in this group of HIV-infected children. The baseline level of CD154 expressed on T helper cells did, however, correlate with the concentration of immunoglobulin A in serum. We conclude that HIV-infected children have impaired regulation of CD154 expression which may contribute to the immune dysregulation commonly observed.","PeriodicalId":10395,"journal":{"name":"Clinical Diagnostic Laboratory Immunology","volume":"156 1","pages":"1104 - 1109"},"PeriodicalIF":0.0,"publicationDate":"2001-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83207520","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-11-01DOI: 10.1128/CDLI.8.6.1044-1048.2001
C. Godshall, A. Lentsch, J. Peyton, M. Scott, W. Cheadle
ABSTRACT The signal transducer and activator of transcription factor 4 (STAT4) pathway mediates the intracellular effects of interleukin-12 (IL-12), leading to the production of gamma interferon, induction of a T helper type 1 response, and increased natural killer cell cytotoxicity. The purpose of this study was to determine the role of the STAT4 pathway during polymicrobial peritonitis in the cecal ligation and puncture (CLP) model. CLP was performed on STAT4-deficient (STAT4−/−) and wild-type control (BALB/c) mice. At 4 h after CLP, STAT4−/− mice had significantly higher bacterial counts in the peritoneal lavage fluid, liver, and blood. This difference persisted for 18 h in the peritoneal lavage fluid and blood. Neutrophil migration to the site of infection and into remote tissues was unaffected. Despite higher bacterial counts locally and systemically, STAT4−/− mice had a lower mortality rate than BALB/c controls. In contrast, blockade of IL-12 in BALB/c mice was detrimental to host survival. A blunted serum IL-12 response at 18 h after CLP was exhibited in STAT4−/− mice. These results suggest several critical roles for the STAT4 pathway in the resolution of polymicrobial infections. Additionally, the disparate effects observed with IL-12 blockade and STAT4 deficiency on host survival suggest that IL-12 may activate alternate pathways promoting survival.
{"title":"STAT4 Is Required for Antibacterial Defense but Enhances Mortality during Polymicrobial Sepsis","authors":"C. Godshall, A. Lentsch, J. Peyton, M. Scott, W. Cheadle","doi":"10.1128/CDLI.8.6.1044-1048.2001","DOIUrl":"https://doi.org/10.1128/CDLI.8.6.1044-1048.2001","url":null,"abstract":"ABSTRACT The signal transducer and activator of transcription factor 4 (STAT4) pathway mediates the intracellular effects of interleukin-12 (IL-12), leading to the production of gamma interferon, induction of a T helper type 1 response, and increased natural killer cell cytotoxicity. The purpose of this study was to determine the role of the STAT4 pathway during polymicrobial peritonitis in the cecal ligation and puncture (CLP) model. CLP was performed on STAT4-deficient (STAT4−/−) and wild-type control (BALB/c) mice. At 4 h after CLP, STAT4−/− mice had significantly higher bacterial counts in the peritoneal lavage fluid, liver, and blood. This difference persisted for 18 h in the peritoneal lavage fluid and blood. Neutrophil migration to the site of infection and into remote tissues was unaffected. Despite higher bacterial counts locally and systemically, STAT4−/− mice had a lower mortality rate than BALB/c controls. In contrast, blockade of IL-12 in BALB/c mice was detrimental to host survival. A blunted serum IL-12 response at 18 h after CLP was exhibited in STAT4−/− mice. These results suggest several critical roles for the STAT4 pathway in the resolution of polymicrobial infections. Additionally, the disparate effects observed with IL-12 blockade and STAT4 deficiency on host survival suggest that IL-12 may activate alternate pathways promoting survival.","PeriodicalId":10395,"journal":{"name":"Clinical Diagnostic Laboratory Immunology","volume":"15 1","pages":"1044 - 1048"},"PeriodicalIF":0.0,"publicationDate":"2001-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83353616","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-11-01DOI: 10.1128/CDLI.8.6.1049-1055.2001
J. Veling, F. V. van Zijderveld, A. M. van Zijderveld-van Bemmel, Y. Schukken, H. Barkema
ABSTRACT Two enzyme-linked immunosorbent assays (ELISAs) for the detectingSalmonella enterica subsp. entericaserovar Dublin antibodies in bulk milk were developed and evaluated for potential use in control programs. The ELISAs were based on either lipopolysacharide (LPS ELISA) or flagellar antigen (GP ELISA). Sensitivity was determined with 79 case herds with a wide range of clinical signs. Specificity was determined with 125 Dutch and 200 Swedish control herds. The relation between antibodies in bulk milk, antibodies in serum, and the level of milk production of individual cows was studied with 61 case herds. The optimal optical density (OD) values of the LPS ELISA and the GP ELISA were determined to be 0.2 and 0.5, respectively. The sensitivities of the LPS ELISA and the GP ELISA were 54 and 63%, respectively, with a specificity of 98% for both ELISAs with samples from the Dutch control herds. The specificities for samples from the Swedish herds were 100% for the LPS ELISA and 95% for the GP ELISA. The sensitivity of the combination of tests was 65% when samples were run in parallel, and the specificity was 100% when samples were run in series, irrespective of whether the samples came from Dutch or Swedish control herds. The variance (R2) in the OD value for bulk milk samples could be explained by the percentage of seropositive lactating cows in a herd with the LPS ELISA for 51% of the samples and with the GP ELISA for 72%. The variance in the OD value was best explained by the combination of the percentage of seropositive lactating cows in the herd and the mean log10 serum antibody titer for that herd (R2 = 62% for the LPS ELISA andR2 = 75% for the GP ELISA). Case herds more often tested negative by the ELISA with bulk milk when the percentage of seropositive lactating cows was less than 5%. It is concluded that both ELISAs with bulk milk can be used in control programs to distinguish between infected and noninfected herds. Specificity can be increased by using the two tests in combination. Sensitivity was relatively low for both single tests and both tests combined.
{"title":"Evaluation of Two Enzyme-Linked Immunosorbent Assays for Detecting Salmonella enterica subsp.enterica Serovar Dublin Antibodies in Bulk Milk","authors":"J. Veling, F. V. van Zijderveld, A. M. van Zijderveld-van Bemmel, Y. Schukken, H. Barkema","doi":"10.1128/CDLI.8.6.1049-1055.2001","DOIUrl":"https://doi.org/10.1128/CDLI.8.6.1049-1055.2001","url":null,"abstract":"ABSTRACT Two enzyme-linked immunosorbent assays (ELISAs) for the detectingSalmonella enterica subsp. entericaserovar Dublin antibodies in bulk milk were developed and evaluated for potential use in control programs. The ELISAs were based on either lipopolysacharide (LPS ELISA) or flagellar antigen (GP ELISA). Sensitivity was determined with 79 case herds with a wide range of clinical signs. Specificity was determined with 125 Dutch and 200 Swedish control herds. The relation between antibodies in bulk milk, antibodies in serum, and the level of milk production of individual cows was studied with 61 case herds. The optimal optical density (OD) values of the LPS ELISA and the GP ELISA were determined to be 0.2 and 0.5, respectively. The sensitivities of the LPS ELISA and the GP ELISA were 54 and 63%, respectively, with a specificity of 98% for both ELISAs with samples from the Dutch control herds. The specificities for samples from the Swedish herds were 100% for the LPS ELISA and 95% for the GP ELISA. The sensitivity of the combination of tests was 65% when samples were run in parallel, and the specificity was 100% when samples were run in series, irrespective of whether the samples came from Dutch or Swedish control herds. The variance (R2) in the OD value for bulk milk samples could be explained by the percentage of seropositive lactating cows in a herd with the LPS ELISA for 51% of the samples and with the GP ELISA for 72%. The variance in the OD value was best explained by the combination of the percentage of seropositive lactating cows in the herd and the mean log10 serum antibody titer for that herd (R2 = 62% for the LPS ELISA andR2 = 75% for the GP ELISA). Case herds more often tested negative by the ELISA with bulk milk when the percentage of seropositive lactating cows was less than 5%. It is concluded that both ELISAs with bulk milk can be used in control programs to distinguish between infected and noninfected herds. Specificity can be increased by using the two tests in combination. Sensitivity was relatively low for both single tests and both tests combined.","PeriodicalId":10395,"journal":{"name":"Clinical Diagnostic Laboratory Immunology","volume":"1 1","pages":"1049 - 1055"},"PeriodicalIF":0.0,"publicationDate":"2001-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89586692","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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