Addition of exogenous cytokines in mixed lymphocyte culture for selecting related donors for bone marrow transplantation.
BACKGROUND
Mixed lymphocyte culturing has led to conflicting opinions regarding the selection of donors for bone marrow transplantation. The association between a positive mixed lymphocyte culture and the development of graft-versus-host disease (GVHD) is unclear. The use of exogenous cytokines in mixed lymphocyte cultures could be an alternative for increasing the sensitivity of culture tests.


OBJECTIVE
To increase the sensitivity of mixed lymphocyte cultures between donor and recipient human leukocyte antigen (HLA) identical siblings, using exogenous cytokines, in order to predict post-transplantation GVHD and/or rejection.


METHODS
Prospective study.


METHODS
Bone Marrow Transplantation Unit, Universidade Estadual de Campinas.


METHODS
Seventeen patients with hematological malignancies and their respective donors selected for bone marrow transplantation procedures.


METHODS
Standard and modified mixed lymphocyte culturing by cytokine supplementation was carried out using donor and recipient cells typed for HLA.


METHODS
Autologous and allogenic responses in mixed lymphocyte cultures after the addition of IL-4 or IL-2.


RESULTS
In comparison with the standard method, average responses in the modified mixed lymphocyte cultures increased by a factor of 2.0 using IL-4 (p < 0.001) and 6.4 using IL-2 (p < 0.001), for autologous donor culture responses. For donor-versus-recipient culture responses, the increase was by a factor of 1.9 using IL-4 (p < 0.001) and 4.1 using IL-2 (p < 0.001). For donor-versus-unrelated culture responses, no significant increase was observed using IL-4, and a mean response inhibition of 20% was observed using IL-2 (p < 0.001). Neither of the cytokines produced a significant difference in the unrelated control versus recipient cell responses.


CONCLUSIONS
IL-4 supplementation was the best for increasing the mixed lymphocyte culture sensitivity. However, IL-4 also increased autologous responses, albeit less intensively than IL-2. Thus, with this loss of specificity we believe that it is not worth modifying the traditional mixed lymphocyte culture method, even with IL-4 addition.
Visentainer
Jeane Eliete Laguila
JE
Universidade Estadual de Campinas, Campinas, São Paulo, Brazil. jelvisentainer@ibest.com.br
Lieber
Sofia Rocha
SR
Persoli
L gia Beatriz Lopes
Lg
Vigorito
Afonso Celso
AC
Aranha
Francisco Jos Penteado
FJ
Souza
C rmino Antonio de
Cr
D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000328:Adult; D016026:Bone Marrow Transplantation / Q000276:immunology*; D005260:Female; D006086:Graft vs Host Disease / Q000276:immunology / Q000517:prevention & control; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D007376:Interleukin-2 / Q000494:pharmacology*; D015847:Interleukin-4 / Q000494:pharmacology*; D007959:Lymphocyte Culture Test, Mixed / Q000379:methods*; D008297:Male; D011446:Prospective Studies; D012680:Sensitivity and Specificity
D000276:Adjuvants, Immunologic; D007376:Interleukin-2; D015847:Interleukin-4; D007371:Interferon-gamma