RESUMO
Objective To compare the growth and reproduction of the promastigotes of Leishmania isolates from various endemic areas of visceral leishmaniasis in China in various culture media, so as to provide experimental evidence for selecting an appropriate medium for the culture of Leishmania. Methods A total of 3 × 105 promastigotes of KS-2, Cy and JIASHI-5 Leishmania isolates were inoculated into 1 mL NNN medium, 1 mL M199 medium supplemented with 20% fetal bovine serum medium, 1 mL M199 medium supplemented with 20% horse serum medium, and 1 mL brain heart infusion medium containing heme, respectively. All media were placed at 22 ℃ under a sterile condition, and the number of promastigotes was counted continuously for 8 days under a microscope. The growth curve was plotted for the three Leishmania isolates. Results The promastigotes of KS-2, Cy and JIASHI-5 Leishmania isolates all grew and reproduced in the NNN medium, the M199 medium supplemented with 20% fetal bovine serum medium, and the M199 medium supplemented with 20% horse serum medium. The number of promastigotes of KS-2, Cy and JIASHI-5 Leishmania isolates was all significantly higher in the NNN medium than in the M199 medium supplemented with 20% fetal bovine serum medium, and the M199 medium supplemented with 20% horse serum medium at various time points of culture (all P values < 0.05), and the number of promastigotes of the KS-2 isolate was all significantly greater than that of the Cy and JIASHI-5 isolates in the NNN medium, the M199 medium supplemented with 20% fetal bovine serum medium, and the M199 medium supplemented with 20% horse serum medium at various time points of culture (all P values < 0.05). In ad dition, the promastigotes of KS-2, Cy and JIASHI-5 Leishmania isolates failed to grow and reproduce in the brain heart infusion medium. Conclusions The growth and reproduction of the promastigotes of various Leishmania isolates from various endemic areas of visceral leishmaniasis in China vary in the same culture medium, and the growth and reproduction of a Leishmania isolate vary in different culture media. The NNN medium best fits for the culture of Leishmania isolates in the endemic areas of visceral leishmaniasis in China.
RESUMO
Objective To analyze the protein abundance differences between two Leishmania infantum strains isolated from different epidemiological types of visceral leishmaniasis in China by comparative proteomics method. Methods Tryptic digests of total proteins were analyzed by using liquid chromatography-tandem mass spectrometry (LC-MS/MS), followed by label-free quantitative differential expression analysis. The MS data were analyzed with MaxQuant software (ver 1.3.0.5) against data base. Results This study resulted in the identification of 4 274 proteins across two strains (JIASHI-5 and SC6) . Of these, 1 219 differentially expressed proteins (ratio ＞ 2.0 or ＜ 0.5, P < 0.05) were identified. Considering the proteins differentially or uniquely expressed in the strains, 550 proteins were only found in the JIASHI-5 strain, and 174 proteins were only found in the SC6 strain. Totally 495 differentially proteins were expressed in the two groups, among which 328 proteins were down-regulated and 167 proteins were up-regulated in SC6 strain. Some of the identified differentially expressed proteins were demonstrated and they involved in energy metabolism, stress response, prolonging the lifetime of the infected host cell and survival and proliferation in virulent strains. Conclusion This study reveals a group of differentially expressed proteins and the related biologic function that may lay the foundation for screening and identification of the key Leishmania molecules relative to pathogenicity.
RESUMO
Objective To compare the growth and reproduction of the promastigotes of Leishmania isolates from various endemic areas of visceral leishmaniasis in China in various culture media, so as to provide experimental evidence for selecting an appropriate medium for the culture of Leishmania. Methods A total of 3 × 105 promastigotes of KS-2, Cy and JIASHI-5 Leishmania isolates were inoculated into 1 mL NNN medium, 1 mL M199 medium supplemented with 20% fetal bovine serum medium, 1 mL M199 medium supplemented with 20% horse serum medium, and 1 mL brain heart infusion medium containing heme, respectively. All media were placed at 22 ℃ under a sterile condition, and the number of promastigotes was counted continuously for 8 days under a microscope. The growth curve was plotted for the three Leishmania isolates. Results The promastigotes of KS-2, Cy and JIASHI-5 Leishmania isolates all grew and reproduced in the NNN medium, the M199 medium supplemented with 20% fetal bovine serum medium, and the M199 medium supplemented with 20% horse serum medium. The number of promastigotes of KS-2, Cy and JIASHI-5 Leishmania isolates was all significantly higher in the NNN medium than in the M199 medium supplemented with 20% fetal bovine serum medium, and the M199 medium supplemented with 20% horse serum medium at various time points of culture (all P values < 0.05), and the number of promastigotes of the KS-2 isolate was all significantly greater than that of the Cy and JIASHI-5 isolates in the NNN medium, the M199 medium supplemented with 20% fetal bovine serum medium, and the M199 medium supplemented with 20% horse serum medium at various time points of culture (all P values < 0.05). In ad dition, the promastigotes of KS-2, Cy and JIASHI-5 Leishmania isolates failed to grow and reproduce in the brain heart infusion medium. Conclusions The growth and reproduction of the promastigotes of various Leishmania isolates from various endemic areas of visceral leishmaniasis in China vary in the same culture medium, and the growth and reproduction of a Leishmania isolate vary in different culture media. The NNN medium best fits for the culture of Leishmania isolates in the endemic areas of visceral leishmaniasis in China.
RESUMO
Objective To analyze the protein abundance differences between two Leishmania infantum strains isolated from different epidemiological types of visceral leishmaniasis in China by comparative proteomics method. Methods Tryptic digests of total proteins were analyzed by using liquid chromatography-tandem mass spectrometry (LC-MS/MS), followed by label-free quantitative differential expression analysis. The MS data were analyzed with MaxQuant software (ver 1.3.0.5) against data base. Results This study resulted in the identification of 4 274 proteins across two strains (JIASHI-5 and SC6) . Of these, 1 219 differentially expressed proteins (ratio ＞ 2.0 or ＜ 0.5, P < 0.05) were identified. Considering the proteins differentially or uniquely expressed in the strains, 550 proteins were only found in the JIASHI-5 strain, and 174 proteins were only found in the SC6 strain. Totally 495 differentially proteins were expressed in the two groups, among which 328 proteins were down-regulated and 167 proteins were up-regulated in SC6 strain. Some of the identified differentially expressed proteins were demonstrated and they involved in energy metabolism, stress response, prolonging the lifetime of the infected host cell and survival and proliferation in virulent strains. Conclusion This study reveals a group of differentially expressed proteins and the related biologic function that may lay the foundation for screening and identification of the key Leishmania molecules relative to pathogenicity.