Quantification of cytotoxic T-cell gene transcripts in human lung transplantation.
BACKGROUND
Differentiating between acute rejection and cytomegalovirus (CMV) infection is one of the major challenges of lung transplantation. The aims of this study were to: (1) quantify the transcription of the cytotoxic T lymphocyte (CTL) effector molecules in the bronchoalveolar lavage (BAL) of lung transplant recipients and (2) evaluate the clinical usefulness of this technique.


METHODS
Sixty-six single-lung, double-lung, or heart-lung transplant patients were prospectively enrolled in the study. BAL was performed either for routine surveillance or for acute graft dysfunction. RNA was extracted from BAL cell pellets and underwent competitive reverse transcription-assisted polymerase chain reaction (RT-PCR) for perforin, granzyme B, granulysin, and Fas ligand. Gene transcript analysis was compared to clinical diagnosis established by conventional methods [BAL microbiological and transbronchial biopsy (TBB) analyses].


RESULTS
After exclusion of several BAL according to the study criteria, 62 BAL were submitted for data analysis. Significantly higher expression of all the analyzed transcripts was found during CMV infection, compared with each of the other defined diagnostic categories, namely nonsignificant pathology, acute rejection, and nonviral pulmonary infection.


CONCLUSIONS
Quantification by competitive RT-PCR of the CTL effector molecule transcripts (perforin, granzyme B, granulysin, and Fas ligand) could represent a valuable tool for the differential diagnosis of graft dysfunction in lung transplantation.
Soccal
P M
PM
Department of Internal Medicine, Stanford University Medical Center, California 94305-5236, USA.
Doyle
R L
RL
Jani
A
A
Chang
S
S
Akindipe
O A
OA
Poirier
C
C
Pavlakis
M
M
D000328:Adult; D000945:Antigens, Differentiation, T-Lymphocyte / Q000235:genetics; D001992:Bronchoalveolar Lavage Fluid / Q000276:immunology; D003586:Cytomegalovirus Infections / Q000276:immunology*; D053222:Fas Ligand Protein; D005260:Female; D006084:Graft Rejection / Q000276:immunology*; D053804:Granzymes; D006801:Humans; D016040:Lung Transplantation / Q000009:adverse effects*; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D011446:Prospective Studies; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000235:genetics; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
D000945:Antigens, Differentiation, T-Lymphocyte; C505661:FASLG protein, human; D053222:Fas Ligand Protein; C075005:GNLY protein, human; D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D012333:RNA, Messenger; D054353:Perforin; C509213:GZMB protein, human; D053804:Granzymes; D012697:Serine Endopeptidases