The frequency of defective genes in vif and vpr genes in 20 hemophiliacs is associated with Korean Red Ginseng and highly active antiretroviral therapy: the impact of lethal mutations in vif and vpr genes on HIV-1 evolution

Abstract

Background: We have reported that internal deletions in the nef, gag, and pol genes in HIV-1-infected patients are induced in those treated with Korean Red Ginseng (KRG). KRG delays the development of resistance mutations to antiretroviral drugs.

Methods: The vif-vpr genes over 26 years in 20 hemophiliacs infected with HIV-1 from a single source were sequenced to investigate whether vif-vpr genes were affected by KRG and KRG plus highly active antiretroviral therapy (ART) (hereafter called GCT) and compared the results with our previous data.

Results: A significantly higher number of in-frame small deletions were found in the vif-vpr genes of KRG-treated patients than at the baseline, in control patients, and in ART-alone patients (p < 0.001). These were significantly reduced in GCT patients (p < 0.05). In contrast, sequences harboring a premature stop codon (SC) were more significant in GCT patients (10.1%) than in KRG-alone patients, control (p < 0.01), and ART-alone patients (p = 0.078 for peripheral blood mononuclear cells). The proportion of SC in Vpr was similar to that in Vif, whereas the proportion of sequences revealing SC in the env-nef genes was significantly lower than that in the pol-vif-vpr genes (p < 0.01). The genetic distance was 1.8 times higher in the sequences harboring SC than in the sequences without SC (p < 0.001). Q135P in the vif gene is significantly associated with rapid progression to AIDS (p < 0.01).

Conclusion: Our data show that KRG might induce sΔ in the vif-vpr genes and that vif-vpr genes are similarly affected by lethal mutations.

Keywords: HIV-1; Korean Red Ginseng; Lethal mutations; Small deletion; vif-vpr genes.

Fig. 1

The proportion of the sequences harboring deletions and in-frame stop codon (SC) in the vif gene. (A) Proportion of patients with internal deletions (IDs), small deletions (SDs), and SCs. Despite the absence of ART in HP 1 and HP 5, all defects were significantly higher in HPs treated with KRG and/or GCT than in the control and ART-alone groups. The proportion of patients with SC was significantly higher in the ART patients than in the control patients. (B) At the sample level, the proportion of amplicon with SC in GCT patients was significantly higher than in the control and KRG patients, and the proportion of SD in KRG patients was significantly higher than in the control, GCT, and even ART-alone patients. (C) SD (≤15-bp) in the vif gene on KRG intake was significantly higher than at baseline and in the control patients (p < 0.001). Its detection was significantly decreased during GCT than on KRG treatment (p < 0.001). (D) The proportion of the sequences harboring SC on GCT was significantly higher than that on KRG treatment and in control patients, whereas it was similar to ART. The proportion was similar among control patients, at baseline, and in KRG patients. ART, antiretroviral therapy; GCT, KRG plus highly active antiretroviral therapy; HP, hemophiliac; KRG, Korean Red Ginseng; PBMC, peripheral blood mononuclear cell.

Fig. 2

Association of Vif Q135P with rapid progression. Q135P was found in Donor O and eight HPs (Fig. S1) and significantly associated with accelerated progression to AIDS. HP, hemophiliac.

Fig. 3

The proportion of the sequences harboring in-frame stop codons in the pol, vif, and vpr genes was significantly and similarly increased during ART, whereas there was no such increase in the env and nef genes [7]. ART, antiretroviral therapy.

Fig. 4

Comparison of genetic distances in the sequences with no stop codons and in-frame stop codons. There were significant difference in the genetic distance between the sequences with no stop codons and in-frame stop codons (2.9 ± 0.8% versus 5.4 ± 1.7%, respectively) over 213 ± 21 and 232 ± 33 months compared with the corresponding earliest sequences from Donors O and P, respectively (p < 0.0001).