Methods and compositions to predict and detect acute rejection
View Patent ↗In some embodiments, a method to detect acute rejection in allograft from is described. In some embodiments, a method to anticipate an episode of acute rejection in allografts is also described. In some embodiments, a kit for detecting or predicting acute transplant rejection of a transplanted organ is described.
1. A method comprising:
obtaining a urinary cell sample from a human patient with a renal allograft but without acute cellular rejection of the allograft;
quantifying mRNA expression levels of perforin, granzyme B, PI-9, IP-10, CD3, FoxP3, and CXCR3 in the urinary cell sample of the patient;
comparing the quantified mRNA expression levels in the urinary cell sample to a corresponding baseline level of mRNA expression of perforin, granzyme B, PI-9, IP-10, CD3, FoxP3, and CXCR3 in urinary cells from a healthy subject or a subject with a non-rejecting renal allograft;
detecting upregulation of mRNA expression of perforin, granzyme B, PI-9, IP-10, CD3, and CXCR3 in the urinary cell sample of the patient and initiating treatment with, or providing a modified dose of, an anti-rejection agent to the patient, or
detecting upregulation of mRNA expression of perforin, IP-10, FoxP3, and CXCR3 in the urinary cell sample of the patient and initiating treatment with, or providing a modified dose of, an anti-rejection agent to the patient; and
obtaining an additional urinary cell sample from the patient after said initiating treatment with, or providing a modified dose of, an anti-rejection agent, and quantifying mRNA expression levels of perforin, granzyme B, PI-9, IP-10, CD3, FoxP3, and CXCR3 in the additional urinary cell sample from the patient.
2. The method according to claim 1 , further comprising comparing log-transformed mRNA expression levels of perforin, granzyme B, PI-9, CD3, FoxP3, and CXCR3 in a urine cell sample from the patient to corresponding log-transformed baseline levels of mRNA expression of perforin, granzyme B, PI-9, IP-10, CD3, FoxP3, and CXCR3 in urine cells from a healthy subject or a subject with a non-rejecting renal allograft.
3. The method according to claim 2 , wherein the log-transformed mRNA levels of perforin, granzyme B, PI-9, IP-10, CD3, FoxP3, and CXCR3 are determined by (i) normalizing mRNA levels to 18S rRNA using a logistic regression model of perforin, granzyme B, PI-9, IP-10, CD3, FoxP3, or CXCR3, or (ii) a weighted combination of at least three log transformed, normalized mRNA levels of perforin, IP-10, FoxP3, granzyme B, PI-9, CD3, or CXCR3 based on a logistic regression model.
4. A method according to claim 1 , further comprising diagnosing future acute rejection when upregulation of mRNA levels is detected in the urine cell sample from the patient.
5. A method according to claim 1 , wherein the method further comprises determining the patient's serum creatinine level in peripheral blood.
6. A method according to claim 1 , wherein the anti-rejection agent is at least one of azathioprine, cyclosporine, FK506, tacrolimus, mycophenolate mofetil, anti-CD25 antibody, antithymocyte globulin, rapamycin, ACE inhibitors, perillyl alcohol, anti-CTLA4 antibody, anti-CD40L antibody, anti-thrombin III, tissue plasminogen activator, antioxidants, anti-CD 154, anti-CD3 antibody, thymoglobin, OKT3, corticosteroid, or a combination thereof.
7. The method of claim 1 , wherein quantifying mRNA expression levels comprises quantifying mRNA levels using one or more probes or primers selected from the group consisting of SEQ ID NO: 1-21, 28, 29 and 30.