Methods of evaluating transplant rejection

A method of detecting acute transplant rejection in a host comprising the steps of:

• a) obtaining a post-transplantation fluid biological sample comprising peripheral blood mononuclear cells containing immune activation marker genes and a constitutively expressed gene from the host, wherein the fluid biological sample is selected from the group consisting of blood and lymphatic fluid;
• b) isolating RNA from the sample thereby obtaining isolated RNA;
• c) obtaining complementary DNA from the isolated RNA;
• d) quantifying cDNA encoding the immune activation marker genes selected from the group consisting of perforin, granzyme B and Fas ligand and the constitutively expressed gene in the sample;
• e) calculating the ratio of the immune activation marker gene cDNA of step d) to the cDNA of the constitutively expressed gene to determine the magnitude of expression of the immune activation marker genes;
• and
• f) determining whether there is a heightened magnitude of expression of any two of the immune activation marker genes as compared to the magnitude of expression of the constitutively expressed gene;
• wherein a heightened magnitude of gene expression of at least two of the immune activation marker genes indicates acute transplant rejection.

A method of screening for acute transplant rejection in a host comprising the steps of:

• a) obtaining a post-transplantation fluid biological sample comprising peripheral blood mononuclear cells containing immune activation marker genes selected from the group consisting of perforin, granzyme B and Fas ligand and constitutively expressed gene from the host, wherein the fluid biological sample is selected from the group consisting of blood and lymphatic fluid;
• b) determining the presence or absence in the sample a gene characteristic of an infectious agent marker;
• and
• c) determining whether there is a heightened gene expression of at least two of said immune activation marker genes, wherein said heightened gene expression in the absence of the infectious agent marker gene indicates acute transplant rejection.

A method of screening for the presence of acute transplant rejection in a host comprising the steps of:

• a) determining the magnitude of expression of at least two immune activation marker genes selected from the group consisting of perforin, granzyme B and Fas ligand genes in a post-transplantation fluid biological sample comprising peripheral blood mononuclear cells from the host by competitive reverse transcription polymerase chain reaction, wherein the
• fluid biological sample is selected from the group consisting of blood and lymphatic fluid;
• and
• b) comparing the magnitude of expression of at least two of said immune activation marker genes in the sample with a normal range of values of the expression of the genes;
• wherein the magnitude of the expression of any two of the genes of step a) that is greater than the magnitude of expression of the normal range of values indicates acute transplant rejection.

A method of assessing the efficacy of anti-rejection therapy during acute transplant rejection in a transplant host comprising the steps of:

• a) quantifying the magnitude of expression of at least two immune activation marker genes selected from the group consisting of perforin, granzyme B, and Fas ligand genes in a first fluid biological sample comprising peripheral blood mononuclear cells selected from the group consisting of blood and lymphatic fluid obtained from the host before administration of the therapy, thereby obtaining a pre-administration quantity of the expression of the transcripts of each gene;
• b) administering anti-rejection therapy to the host;
• c) quantifying the magnitude of expression of the genes in a second fluid biological sample comprising peripheral blood mononuclear cells selected from the group consisting of blood and lymphatic fluid obtained from the host after administration of the therapy, thereby obtaining a post-administration quantity for expression of the transcripts of each of the genes;
• and
• d) comparing the pre-administration quantity of the transcripts of each of said genes to the post-administration quantity of the transcripts of the same gene, wherein the post-administration magnitude of the transcripts of any two of said genes that is less than the pre-administration magnitude of the transcripts of the same two genes indicate the efficacy of the anti-rejection therapy during acute transplant rejection.