Objective:To study the genetic diversity at the msp—1,msp—2,and glurp genes of Plasmodium falciparum(P.falciparum) isolates from 3 endemic areas in Thailand:Tak.Kanchanaburi and Ranong provinces.Methods:A total of 1...Objective:To study the genetic diversity at the msp—1,msp—2,and glurp genes of Plasmodium falciparum(P.falciparum) isolates from 3 endemic areas in Thailand:Tak.Kanchanaburi and Ranong provinces.Methods:A total of 144 P.falciparum isolates collected prior to Irealmenl during January.2012 to June,2013 were genotyped.DNA was extracted:allele frequency and diversity of msp-1.msp-2,and glurp genes were investigated by nested polymerase chain reaction.Results:P.falciparum isolates in this study had high rate of multiple genotypes infection(96.5%)with an overall mean multiplicity of infection of 3.21.The distribution of allelic families of msp-1was significantly different among isolales from Tak.kanchanahnri.and Ranong but not for the msp-2.K1 and MAD20 were the predominant allelic families at the msp-1 gene,whereas alleles belonging to 3D7 were more frequent at the msp-2 gene.The glurp gene had the least diverse alleles.Population structure of P.falciparum isolates from Tak and Ranong was quite similar as revealed by the presence of similar proportions of MAD20 and K1 alleles at msp-1 loci.3D7 and FC27 alleles at msp-2 loci as well as comparable mean MOI.Isolates from Kanchanaburi had different structures;the most prevalent alleles were K1 and RO33.Conclusions:The present study shows that P.falciparum isolates from Tak and Ranong provinces had similar allelic pattern of msp—1 and msp—2 and diversity but different from Kanchanaburi isolates.These allelic variant profiles are valuable baseline data for future epidemiological study of malaria transmission and for continued monitoring of polymorphisms associated with antimalarial drug resistance in these areas.展开更多
Objective: To determine the effects of toll-like receptor 2(TLR-2) agonist, Pam3 CSK4, on cellular and humoral immune response against recombinant Mycobacterium bovis bacille Calmette-Guérin(rBCG) expressing the ...Objective: To determine the effects of toll-like receptor 2(TLR-2) agonist, Pam3 CSK4, on cellular and humoral immune response against recombinant Mycobacterium bovis bacille Calmette-Guérin(rBCG) expressing the C-terminus of merozoite surface protein-1 of Plasmodium falciparum.Methods: Six groups of mice(n=6 per group) received intraperitoneal phosphate buffered saline T80(PBS-T80), BCG or rBCG in the presence or absence of Pam3 CSK4. Enzymelinked immunosorbent assay was carried out to measure serum total IgG, IgG1, IgG2 a, and Ig G2 b production. Spleens were also harvested and splenocytes were co-cultured with rBCG antigen for in vitro determination of IL-4 and IFN-γ via enzyme-linked immunosorbent assay.Results: The production of total IgG and the isotype IgG1, IgG2 a and IgG2 b was significantly higher in rBCG-immunised mice than in the BCG and PBS-T80-immunised mice, and Pam3 CSK4 further enhanced their productions. A similar pattern was also observed in IFN-γ production. Moreover, there was no significant difference in IL-4 production in all groups either in the presence or absence of Pam3 CSK4.Conclusions: We present evidence of the adjuvant effects of TLR-2 agonist in enhancing the production of total IgG, IgG1, IgG2 a, IgG2 b, as well as IFN-γ in response to rBCG. However, the presence or absence of Pam3 CSK4 had no effect on IL-4 production.展开更多
Objective: To determine the role of toll-like receptor 4(TLR-4) in eliciting cellular and humoral immune responses against recombinant Mycobacterium bovis bacille Calmette-Guérin(rB CG) expressing the C-terminus ...Objective: To determine the role of toll-like receptor 4(TLR-4) in eliciting cellular and humoral immune responses against recombinant Mycobacterium bovis bacille Calmette-Guérin(rB CG) expressing the C-terminus of merozoite surface protein-1 of Plasmodium falciparum. Methods: Six groups of mice(n=6 per group) were injected with phosphate buffered saline T80, BCG or r BCG intraperitoneally, in the presence or absence of a TLR-4 inhibitor; TAK-242. Enzyme-linked immunosorbent assay was carried out for serum total IgG, IgG 1, Ig G2 a and Ig G2 b determination. Spleens were also harvested and splenocytes cultured for determination of intracellular cytokines; IL-4 and IFN-毭 via enzyme-linked immunosorbent assay. Results: The production of total Ig G, and the subclasses Ig G1, Ig G2 a and Ig G2 b was significantly higher in rB CG-immunised mice than BCG and phosphate buffered saline immunised mice in the absence of TAK-242. A significant rise in total IgG occurred with more booster immunisations. The level of IgG 2 a was highest, followed by IgG 2 b, then IgG 1. The production of both IL-4 and IFN-were inhibited in t毭 was also highest in the rB CG immunised groups. These significant riseshe presence of TAK-242. Conclusions: We present evidence of the role of TLR-4 in the increased production of total IgG, IgG 1, IgG 2 a and IgG 2 b, as well as IL-4 and IFN-毭 in response to our rB CG construct.展开更多
基金Supported by a grant from Research Institute,Bansomdejchaopraya Rajabhat University(Grant no.2555)
文摘Objective:To study the genetic diversity at the msp—1,msp—2,and glurp genes of Plasmodium falciparum(P.falciparum) isolates from 3 endemic areas in Thailand:Tak.Kanchanaburi and Ranong provinces.Methods:A total of 144 P.falciparum isolates collected prior to Irealmenl during January.2012 to June,2013 were genotyped.DNA was extracted:allele frequency and diversity of msp-1.msp-2,and glurp genes were investigated by nested polymerase chain reaction.Results:P.falciparum isolates in this study had high rate of multiple genotypes infection(96.5%)with an overall mean multiplicity of infection of 3.21.The distribution of allelic families of msp-1was significantly different among isolales from Tak.kanchanahnri.and Ranong but not for the msp-2.K1 and MAD20 were the predominant allelic families at the msp-1 gene,whereas alleles belonging to 3D7 were more frequent at the msp-2 gene.The glurp gene had the least diverse alleles.Population structure of P.falciparum isolates from Tak and Ranong was quite similar as revealed by the presence of similar proportions of MAD20 and K1 alleles at msp-1 loci.3D7 and FC27 alleles at msp-2 loci as well as comparable mean MOI.Isolates from Kanchanaburi had different structures;the most prevalent alleles were K1 and RO33.Conclusions:The present study shows that P.falciparum isolates from Tak and Ranong provinces had similar allelic pattern of msp—1 and msp—2 and diversity but different from Kanchanaburi isolates.These allelic variant profiles are valuable baseline data for future epidemiological study of malaria transmission and for continued monitoring of polymorphisms associated with antimalarial drug resistance in these areas.
基金supported by the Universiti Sains Malaysia(USM)Research University(RU)Grants(No.1001/PPSK/8011100)
文摘Objective: To determine the effects of toll-like receptor 2(TLR-2) agonist, Pam3 CSK4, on cellular and humoral immune response against recombinant Mycobacterium bovis bacille Calmette-Guérin(rBCG) expressing the C-terminus of merozoite surface protein-1 of Plasmodium falciparum.Methods: Six groups of mice(n=6 per group) received intraperitoneal phosphate buffered saline T80(PBS-T80), BCG or rBCG in the presence or absence of Pam3 CSK4. Enzymelinked immunosorbent assay was carried out to measure serum total IgG, IgG1, IgG2 a, and Ig G2 b production. Spleens were also harvested and splenocytes were co-cultured with rBCG antigen for in vitro determination of IL-4 and IFN-γ via enzyme-linked immunosorbent assay.Results: The production of total IgG and the isotype IgG1, IgG2 a and IgG2 b was significantly higher in rBCG-immunised mice than in the BCG and PBS-T80-immunised mice, and Pam3 CSK4 further enhanced their productions. A similar pattern was also observed in IFN-γ production. Moreover, there was no significant difference in IL-4 production in all groups either in the presence or absence of Pam3 CSK4.Conclusions: We present evidence of the adjuvant effects of TLR-2 agonist in enhancing the production of total IgG, IgG1, IgG2 a, IgG2 b, as well as IFN-γ in response to rBCG. However, the presence or absence of Pam3 CSK4 had no effect on IL-4 production.
基金supported by the Universiti Sains Malaysia(USM) Fundamental Research Grant Scheme(FRGS)(No.203/PPSK/6171158)
文摘Objective: To determine the role of toll-like receptor 4(TLR-4) in eliciting cellular and humoral immune responses against recombinant Mycobacterium bovis bacille Calmette-Guérin(rB CG) expressing the C-terminus of merozoite surface protein-1 of Plasmodium falciparum. Methods: Six groups of mice(n=6 per group) were injected with phosphate buffered saline T80, BCG or r BCG intraperitoneally, in the presence or absence of a TLR-4 inhibitor; TAK-242. Enzyme-linked immunosorbent assay was carried out for serum total IgG, IgG 1, Ig G2 a and Ig G2 b determination. Spleens were also harvested and splenocytes cultured for determination of intracellular cytokines; IL-4 and IFN-毭 via enzyme-linked immunosorbent assay. Results: The production of total Ig G, and the subclasses Ig G1, Ig G2 a and Ig G2 b was significantly higher in rB CG-immunised mice than BCG and phosphate buffered saline immunised mice in the absence of TAK-242. A significant rise in total IgG occurred with more booster immunisations. The level of IgG 2 a was highest, followed by IgG 2 b, then IgG 1. The production of both IL-4 and IFN-were inhibited in t毭 was also highest in the rB CG immunised groups. These significant riseshe presence of TAK-242. Conclusions: We present evidence of the role of TLR-4 in the increased production of total IgG, IgG 1, IgG 2 a and IgG 2 b, as well as IL-4 and IFN-毭 in response to our rB CG construct.