Burkholderia pseudomallei proteins presented by monocyte-derived dendritic cells stimulate human memory T cells in vitro.
Melioidosis is a severe infectious disease caused by the saprophytic facultative intracellular pathogen Burkholderia pseudomallei. The disease is endemic in Southeast Asia and Northern Australia, and no effective vaccine exists. To describe human cell-mediated immune responses to B. pseudomallei and to identify candidate antigens for vaccine development, the ability of antigen-pulsed monocyte-derived dendritic cells (moDCs) to trigger autologous T-cell responses to B. pseudomallei and its products was tested. moDCs were prepared from healthy individuals exposed or not exposed to B. pseudomallei, based on serological evidence. These were pulsed with heat-killed B. pseudomallei or purified antigens, including ABC transporters (LolC, OppA, and PotF), Bsa type III secreted proteins (BipD and BopE), tandem repeat sequence-containing proteins (Rp1 and Rp2), flagellin, and heat shock proteins (Hsp60 and Hsp70), prior to being mixed with autologous T-cell populations. After pulsing of cells with either heat-killed B. pseudomallei, LolC, or Rp2, coculturing the antigen-pulsed moDCs with T cells elicited gamma interferon production from CD4(+) T cells from seropositive donors at levels greater than those for seronegative donors. These antigens also induced granzyme B (cytotoxic) responses from CD8(+) T cells. Activation of antigen-specific CD4(+) T cells required direct contact with moDCs and was therefore not dependent on soluble mediators. Rp peptide epitopes recognized by T cells in healthy individuals were identified. Our study provides valuable novel data on the induction of human cell-mediated immune responses to B. pseudomallei and its protein antigens that may be exploited in the rational development of vaccines to combat melioidosis.
The Centre for Research & Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand.
Patcharaporn
P
Tippayawat
Maneerat
M
Pinsiri
Darawan
D
Rinchai
Donporn
D
Riyapa
Amornrat
A
Romphruk
Yunn-Hwen
YH
Gan
Raymond L
RL
Houghton
Philip L
PL
Felgner
Richard W
RW
Titball
Mark P
MP
Stevens
Edouard E
EE
Galyov
Gregory J
GJ
Bancroft
Ganjana
G
Lertmemongkolchai
D001426:Bacterial Proteins / Q000378:metabolism*; D016957:Burkholderia pseudomallei / Q000276:immunology* / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000502:physiology*; D003713:Dendritic Cells / Q000378:metabolism*; D019353:Endemic Diseases; D018984:Epitopes, T-Lymphocyte; D015964:Gene Expression Regulation, Bacterial; D006358:Hot Temperature; D006801:Humans; D008554:Melioidosis / Q000453:epidemiology / Q000276:immunology* / Q000382:microbiology*; D057057:Serine Proteases; D020080:Tandem Repeat Sequences
D001426:Bacterial Proteins; D018984:Epitopes, T-Lymphocyte; D057057:Serine Proteases