Study On The Immune Function And Retinal Microvascular Disease Of Kangaibaosheng Pill Combined With HAART On HIV Infection

Objective:To determine TRECs level in CD45RA~+T lymphocytes by real-time quantitative PCR and then to evaluate the recent thymic output function,and to evaluate the effectiveness of Kangaibaosheng pill combined with HAART in improving immune function via regular detection and assessment of TCR V?gene profile,T lymphocyte count,CD4~+CD45RA~+and CD4~+CD45RO~+T lymphocyte counts,plasma viral load changes,immune reconstitution rate,and retinal microvascular changes in HIV-infected patients,providing the data support for the clinical application of Kangaibaosheng pill,a new evidence for the optimization of clinical immune reconstitution regimen after HAART,and the basic foundation for the clinical application of integrated traditional Chinese and Western medicine in the treatment of HIV infected persons.Methods:Three hundred HIV-infected patients who met the inclusion criteria were randomly divided into 2 groups,namely the HAART group,and the HAART+Kangaibaosheng pill group,with 150 cases each.General information,content of TRECs,TCR V?gene profile,T lymphocyte count,CD4~+CD45RA~+and CD4~+CD45RO~+T lymphocyte counts,plasma viral load changes,immune reconstitution rate,and retinal microvascular changes(with 40 cases each)were dynamically monitored and evaluated at 0 and 6 months of treatment(Month 0 and Month 6),specifically:(1)Effect of HAART+Kangaibaosheng pill on T lymphocytes(cell counts of CD4~+T,CD8~+T,and CD3~+T,and CD4/CD8 ratio)and CD4~+CD45RA~+and CD4~+CD45RO~+T lymphocyte counts in HIV-infected patients was analyzed by BD FACSCalibur flow cytometry;(2)Effect of HAART+Kangaibaosheng pill on TRECs content in AIDS patients was determined by using real-time quantitative PCR to reflect the recent thymic output function;(3)Effect of HAART+Kangaibaosheng pill on the TCR V?gene profile in HIV infected persons was analyzed by RT-PCR;(4)Effect of HAART~+Kangaibaosheng pill on the plasma viral load in HIV infected persons was analyzed with NucliSENS easyQ HIV-1 virus load testing system;(5)Immune reconstitution rate in the two groups was measured and compared at the end of 6 months of treatment.(6)Retinal microvascular lesions in patients of each group were observed by nonmydriatic fundus camera and retinal image analysis technology to investigate the effect of HAART+Kangaibaosheng pill on the retinal microvascular changes in HIV infected persons.Results:1.Effect of Kangaibaosheng pill combined with HAART on the immune reconstitution in HIV-infected patients(1)Adverse reaction was the main reason for patient drop-out in both groups,with the highest dropout rates of 42.55%and 29.79%,respectively,and the rate of the HAART group was higher than that of the HAART+Kangaibaosheng pill group.(2)Drug adherence index of the HAART group was higher than that of the HAART~+Kangaibaosheng pill group(79.22%and 61.38%,respectively).(3)Patients in the two groups were mainly the young and middle-aged,with more male than female and more patients with Han nationalities than the minorities.The majority of the patients were married with middle school education.Sexual contact and intravenous drug abuse were the main routes of infection.(4)Total symptom and sign score and Karovsky score:at Month 6,total symptom and sign score of the HAART group was significantly increased compared with that of Month 0(P<0.001),and the total score of the HAART+Kangaibaosheng pill group was decreased,but the difference was not significant(P>0.05).The total symptom and sign score of the HAART+Kangaibaosheng pill group was decreased compared with the HAART group with a significant difference(P<0.001).At Month 6,Karovsky score of the HAART group was significantly reduced(P<0.001)and that of the HAART~+Kangaibaosheng pill group was significantly increased(P<0.001)compared with that of Month 0;the Karovsky score of the HAART+Kangaibaosheng pill group was increased compared with the HAART group and the difference was statistically significant(P<0.001).(5)Absolute cell count of CD3~+T lymphocytes:CD3~+T lymphocyte count in the HAART group at Month 6 was not significantly different from that of Month 0(P>0.05),and the value of the HAART+Kangaibaosheng pill group was not significantly different when compared with that of Month 0 and the HAART group,respectively(P>0.05).(6)Absolute cell count of CD4~+T lymphocytes:the cell counts of the HAART group and the HAART+Kangaibaosheng pill group at Month 6 were increased compared with that of Month 0,respectively,with significant differences(P<0.001),and comparison between these two groups revealed a significant difference(P<0.001).(7)Relative cell count of CD4~+T lymphocytes:the cell counts of the HAART group and the HAART+Kangaibaosheng pill group at Month 6 were increased compared with that of Month 0,respectively,with significant differences(P<0.001),and comparison between these two groups revealed a significant difference(P<0.01).(8)Absolute cell count of CD8~+T lymphocytes:compared with Month 0,the cell counts of the HAART group and the HAART+Kangaibaosheng pill group were decreased at Month 6,respectively,with significant differences(P<0.01 and P<0.001),and no significant difference was observed when these two groups were compared(P>0.05).(9)Relative cell count of CD8~+T lymphocytes:compared with Month 0,the cell counts of the HAART group and the HAART+Kangaibaosheng pill group were decreased at Month 6,respectively,with significant differences(P<0.01),and no significant difference was observed when these two groups were compared(P>0.05).(10)CD4/CD8 ratio:CD4/CD8 ratio of the HAART group and the HAART~+Kangaibaosheng pill group were significantly increased at Month 6 compared with that of Month 0(P<0.01 and P<0.001),and the ratio in the HAART+Kangaibaosheng pill group was higher than that of the HAART group with a significant difference(P<0.01).(11)CD4~+CD45RA~+and CD4~+CD45RO~+T lymphocyte counts:compared with Month 0,the cell counts of the CD4~+CD45RA~+and CD4~+CD45RO~+T in the HAART group and the HAART+Kangaibaosheng pill group were significantly increased at Month6(P<0.01 and P<0.001);the CD4~+CD45RA+,CD4~+CD45RO~+T in the HAART~+Kangaibaosheng pill group was higher than that of the HAART group with significant differences(P<0.01).(12)Changes of TRECs content in CD4~+and CD8~+T lymphocytes:TRECs contents in CD4~+and CD8~+T lymphocytes in the HAART group and the HAART+Kangaibaosheng pill group were significantly increased at Month 6 compared with that of Month 0(P<0.01)and compared with the HAART group,the HAART+Kangaibaosheng pill group were significantly increased(P<0.05).(13)Gene profiling of TCR V?family:D values of nine families of V?-2,4,5.2,11,17,20,21,23 and 24 in CD4~+and CD8~+T cells in the two groups after treatment were significantly reduced compared with that before treatment(P<0.05),in which the decrease of D values of the nine families in the HAART+Kangaibaosheng pill group was more prominent than the HAART group and the differences were of statistical significance(P<0.05).(14)Fluctuation of plasma viral load:when six months of treatment was completed,there were no significant differences in the number of cases with plasma viral load<100copy/m L and the percentages of patients with fluctuated viral load between the two groups(P>0.05).(15)Comparison of the immune reconstruction efficiency:immune reconstruction rate in the HAART~+Kangaibaosheng pill group(77.52%)was higher than that of the HAART group(58.87%)and the difference was significant(P<0.05).2.Effect of Kangaibaosheng pill combined with HAART on retinal microvascular pathological changes in patients with AIDS(1)Fundus lesions:no obvious pathological changes were observed before and after treatment for the patients in the two groups.(2)Retinal artery diameter:There were no significant differences in mean artery diameter and artery diameters of the superior and inferior temporal arteries and the superior and inferior nasal arteries between the HAART group and the HAART+Kangaibaosheng pill group before and after 6 months of treatment(P>0.05).(3)Retinal vein diameter:No significant differences was noted in mean vein diameter and vein diameters of the superior and inferior temporal veins and the superior and inferior nasal veins between the HAART group and the HAART+Kangaibaosheng pill group before treatment(P>0.05).After 6 months of treatment,diameter of the superior temporal vein in the patients of the HAART+Kangaibaosheng pill group was significantly decreased compared than that of the HAART group(P<0.05),between the two groups,no significant differences were found in mean vein diameter and the superior nasal vein,the inferior nasal vein,and the inferior temporal vein diameters(P>0.05).(4)Artery-to-vein diameter ratios of retinal arteries and veins:there were no significant differences in the mean artery-to-vein diameter ratio and the artery-to-vein diameter ratios of the superior and inferior temporal arteries and veins and the superior and inferior nasal arteries and veins(P>0.05).Conclusions:1.Effect of Kangaibaosheng pill combined with HAART on the immune reconstitution in HIV-infected patients(1)Adverse reaction is the main cause for the highest dropout rates in both groups.(2)Characteristics of the patients enrolled in the study are in accord with the AIDS epidemiologic features in Yunnan Province.(3)Drug adherence in the patients treated with HAART alone is higher than that of the patients received Kangaibaosheng pill combined with HAART.(4)Kangaibaosheng pill combined with HAART more effectively reduces the clinical symptom and sign of HIV-infected patients and improves quality of life of HIV-infected patients than HAART treatment alone.(5)HAART and Kangaibaosheng pill may not play a therapeutic role by increasing the CD3~+and CD8~+T lymphocyte counts.(6)Kangaibaosheng pill combined with HAART can more effectively increase the CD4~+T lymphocyte count in HIV-infected patients than HAART treatment alone.(7)Although Kangaibaosheng pill combined with HAART significantly increases the CD4/CD8 ratio in HIV-infected patients than HAART treatment alone,the overall ratio of CD4/CD8 is still inverted.(8)Despite that Kangaibaosheng pill combined with HAART can improve the absolute CD4~+CD45RA~+T lymphocyte counts in HIV-infected patients,whether its function is recovered to normal is not determined and it is likely that CD4~+CD45RA~+T lymphocytes in peripheral blood are detected instead of the neonatal CD4~+CD45RA~+T lymphocytes at this moment.(9)Kangaibaosheng pill combined with HAART treatment can better improve the CD45RO~+T cell counts in patients.However,due to the fact that HIV virus may hide in CD4~+CD45RO~+T cells,the increase is of beneficial of harmful effect on the treatment of AIDS is inconclusive.(10)Kangaibaosheng pill combined with HAART can preferably increase the contents of TRECs/10~6 CD4~+T and TRECs/10~6 CD8~+T in HIV-infected patients,whether this increase is induced through the thymus-dependent pathway or thymus-independent pathway,however,still needs to be determined with prolonged treatment time later on.(11)Kangaibaosheng pill combined with HAART treatment can more effectively improve the oligoclonal expression of some TCRV?genes in CD4~+and CD8~+T cells.(12)Mechanism of Kangaibaosheng pill on HIV-infected patients is not related to the inhibition of plasma viral load.(13)Kangaibaosheng pill combined with HAART more significantly improves the success rate of immune reconstitution than the HAART treatment alone.2.Effect of Kangaibaosheng pill combined with HAART on retinal microvascular pathological changes in patients with AIDS(1)No obvious ocular lesions are observed during treatment in patients received HAART alone or Kangaibaosheng pill combined with HAART.(2)Neither HAART alone nor Kangaibaosheng pill combined with HAART causes or aggravates the stenosis of mean artery diameter and diameters of the superior and inferior temporal arteries and the superior and inferior nasal arteries.(3)Kangaibaosheng pill combined with HAART reduces the degree of superior temporal vein dilation.(4)Neither HAART alone nor Kangaibaosheng pill combined with HAART alters the artery-to-vein diameter ratios of the superior and inferior temporal arteries and veins and the superior and inferior nasal arteries and veins.