HIV infection and AIDS are endemic in many malnourished populations. A balanced nourishing diet is fundamental for healthiness and survival for all individuals, including for HIV-infected individuals. Tuberculosis and diarrhea, which occur in many HIV infected individuals, cause by themselves appetite loss, atrophy and weight loss. In HIV-infected individuals, the body energy requirements are higher than in non-HIV infected individuals12345. Following HIV infection, before and after the onset of AIDS, including in AIDS patients receiving highly active antiretroviral treatment (HAART), the energy requirements needed to preserve the body weight increase by 20% to 30%678. In view of the above, the World Health Organization has recommended the inclusion of micronutrients (MMN) administration in any treatment protocol for HIV-infected individuals at any stage of their disease, including during pregnancy and lactation and for children6.

Almost in all randomized controlled trials that studied the effects of MMN supplementation found increased CD4+ T-cell counts or reduced mortality in the group of HIV-infected persons receiving MMN as compared to the HIV-infected persons receiving placebo9101112131415. Different MMN interventions have been evaluated in the various trials conducted and the conclusion from all these studies is that MMN supplementation confers clear clinical benefits to HIV-infected individuals, including to pregnant women and their offspring, regardless of their clinical stage and use of Antiretroviral Therapy (ART)15.

Phytochemicals, chemical compounds that occur naturally in plants, serve as micronutrients. Importantly, some phytochemicals also have additional important beneficial properties, as demonstrated in several clinical studies. For example, some phytochemicals possess radical scavenging activities16 some stimulate nonspecific immunity17 some down regulate inflammatory diseases18 and some have anti-hepatotoxic, anti-lithic, anti-hypertensive, and anti-hepatitis properties19. Interestingly, some phytochemicals demonstrate potent anti-HIV in vitro activity, especially against the HIV-1 protease and integrase, and against gp41 acting as entry inhibitors1619202122232425262728.

Phyto V7 is a specific mix of phytochemicals that has been found to have immune-stimulating properties. This was demonstrated in two separate studies. In the first one, administration of Phyto V7 to chicks vaccinated against Newcastle Disease Virus resulted in enhanced humoral immune responses against the virus 29. In the second study, administration of Phyto V7 to women, infected with Human Papilloma Virus (HPV) and with preneoplastic cervical lesions, resulted in enhanced cervical in situ cellular immune responses and increased clearance of HPV30.

In the current study we studied the clinical and immunological effects of the administration of Phyto V7 on HIV-1 seropositive individuals and AIDS patients, in order to determine if this phytochemical complex may be an important nutritional component to be given to these populations.

Enrollment took place between December 2009 and February 2010 in Hospital de Clínicas, Universidad Nacional de Córdoba, city of Cordoba, Argentina. Clinical interview, anthropometric and hematological analysis were performed prior to enrollment. Only diagnosed HIV-1 seropositive patients with less than 350 CD4+ T-cells per mm^³, who had never received ART, were enrolled. Individuals with co-current infections or positive to hepatitis antibodies were excluded. Written informed consent was obtained from all study participants after explaining the trial aims and specifics in detail. Randomization to each of the detailed groups above was guided by the expressed will of the patient. A total of 28 patients were enrolled and divided into 4 groups as detailed in Figure 1: Control Group 1 (CG1; no Phyto V7, no ART); Control Group 2 (CG2; no Phyto V7, HAART); Test Group 1 (TG1; Phyto V7, no ART); and Test Group 2 (TG2; Phyto V7, HAART). The mean and age range, sex, weight and other characteristics of the enrolled patients per group are detailed in Table 1. The concomitant infections and HAART regimens of each of the patients are detailed in Table 2. In the Test Groups 1 and 2, the patients were requested to consume 2 tablets of Phyto V7 every 8 hours daily for the duration of the trial. Each Phyto V7 tablet contained 760 mg of the following phytochemicals: flavonols (Kaempferol, Quercetin), flavones (Apigenin, Luteolin), hydroxy-cinnamic acids (ferrulic acid), carotenoids (Lutein, Lycopene, Beta carotenne) andorganosulfur compounds, all from vegetal origin31. Cross-sectional clinical and laboratory studies were conducted every 3 months. The CD4 lymphocyte count was measured by conventional flow cytometry. Plasma HIV-1 RNA was measured using an ultraquantitative polymerase chain reaction assay with a lower limit of quantification of 50 copies/mL.

This study demonstrates that the oral administration of a phytochemical complex (Phyto V7) is beneficial to HIV-1 seropositive individuals and AIDS patients, whether receiving antiretroviral therapy or not. There were quantitative improvements in the patients receiving Phyto V7 in their weight, BMI and CD4+ T-cell counts. While there was a mean decrease in the weight and BMI of the patients that did not receive any treatment during the trial, in the patients that received only Phyto V7 there was a significant increase in the weight and BMI both after 3 and 6 months of the study. Interestingly, in the patients that received only ARV treatment, there was a statistical significant increase in weight and BMI only after 6 months of treatment, as compared to the patients that did not receive any treatment. The most notorious increases in weight and BMI were noted in the patients that received both ARV treatment and Phyto V7. Meaningfully, there was a clear statistical difference between the increase in weight and BMI in those patients that received ARV treatment only and those that received the ARV and also the phytochemicals both at 3 and 6 months (Figure 2 and Figure 3), indicating the significant contribution of the phytochemicals to the well-being of the patients. Outstandingly, there was also clear improvement in the CD4+ T-cell counts of the group of patients that received Phyto V7 only as compared to those that did not receive any treatment (Figure 5), which was similar to the increase in CD4+ T-cell counts that occurred in the group of patients that received antiretroviral treatment only. The medical improvement in the patients receiving Phyto V7 was very noticeable according to the treating doctor’s impression and the patient’s feedback (appetite, reduction of diarrhea). While it has been reported that some phytochemicals possess potent anti-HIV in vitro activity1619202122232425262728, apparently their beneficial effects cannot be explained by their direct activity on the virus, as there were no decreases in viremia in the patients that received the phytochemicals only. The results of this study are in accordance with the dramatic improvement in the physical status of a small cohort of 9 terminal AIDS patients following 45 days of administration of Phyto V732 and with the significant improvement in the well-being of 199 HIV-1 infected individuals, who were not undergoing antiretroviral treatment and were recruited as part of the Uruguay National Program of AIDS, which received only a daily administration of Phyto V7 for a period of 90 consecutive days31.

The HAART regimens used in this trial were based on the recommended Argentinian Ministry of Health guidelines at the time the trial was conducted. Those regimens as well as other HAART regimens, when taken with good compliance, can effectively reduce the viral loads to undetectable levels and significantly improve prognosis and well-being of the patients, as indeed was the case in the group of patients in this study that only received the antiretroviral therapy. However, HAART may cause also significant adverse effects, such as lipodystrophy, dyslipidaemia, cardiovascular complications, central and peripheral nervous system disturbances and insulin resistance33343536. HAART may be very problematic to certain populations, such as pregnant women and children3738. The large number of pills needed for HAART also leads to significant problems of compliance and development of resistant virus39. HAART is not implemented in many resource-limited regions where the AIDS epidemic is rampant due to its high cost. Furthermore, the spectrum of adverse effects related to HAART in developing countries may be even more deleterious and hard to treat because of the high prevalence of conditions such as anemia, malnutrition, and tuberculosis and frequent initial presentation with advanced HIV disease40. Taken together, there is a rational for postponing the administration of HAART to HIV-1 infected patients as much as possible, but not too late before the CD4+ T-cell counts are too low 41.

As indicated in this study, in patients that did not receive antiretroviral treatment, the administration of Phyto V7 resulted in an increase in their CD4+ T-cell counts, weight and BMI, indicating that supplementation of this phytochemical mix may improve the capacity of HIV-1 infected individuals to cope with the viral infection and potentially delaying the need to treat them with HAART, postponing the potential complications associated with HAART treatment.

HAART treatment taken with good compliance in most cases results in viremia suppression, immune reconstitution, and reduction in incidence and severity of opportunistic diseases and death42. Similar results were obtained in this study. However, in many HIV-1 infected individuals and AIDS patients undergoing HAART treatment, there is no reconstitution of their immune system, and the CD4+ T-cell counts do not increase even if full plasma viral load suppression is achieved43. This may be further exasperated in AIDS/tuberculosis patients, AIDS patients that already suffer from opportunistic infections, or are co-infected with other parasites, such as helminthes or other viral infections, as is the case in many African countries. It would be very significant if in these individuals the supplementation with phytochemicals would increase their CD4+ T-cell counts. This would be a much simpler associated safe treatment than immune therapies being explored today like cytokine therapies, therapeutic immunization, monoclonal antibodies, immune-modulating drugs, nanotechnology-based approaches and radio immune therapy 4445.

What are the mechanisms by which the phytochemicals benefit HIV-1 seropositive and AIDS patients is not yet clear and should be elucidated. While it has been reported that some phytochemicals possess potent anti-HIV in vitro activity1619202122232425262728, apparently their beneficial effects cannot be explained by their direct activity on the virus, as there were no decreases in viremia in the patients that received the phytochemicals only (Figure 4). In the HAART and PHT co-treatment group (TG2), when looking at the mean viremia of the group (Figure 4), the impression that PHT counteracts the effect of HAART could be reached. However, as explained in the Results section, in the TG2 group there were 6 individuals, all of which had high viral loads at the onset of the trial. Following the HAART treatment and PHT co-treatment in 5 out of the 6 patients, no viral load was detected at the 3 months period (less than ln 3.9, i.e. less than 50 viral RNA copies/ml). Unfortunately, in one individual the viremia remained very high (62685 viral RNA copies/ml) at 3 months examination. It turned out that this particular patient had a resistant virus and only after changing the antiretroviral medication, from the 3rd month of the commencement of the Trial and onward, the HAART treatment was efficacious. This indicates that the PHT co-treatment does not interfere with antiviral activity of HARRT therapy.

Part of their positive effect can be explained as serving as micronutrients, having radical scavenging activities16, stimulating nonspecific immunity17, and by down regulating inflammatory responses18. Importantly, administration of Phyto V7 to 33 women infected with Human Papilloma Virus (HPV) and with preneoplastic cervical lesions, resulted in enhanced cervical in situ cellular immune responses and increased clearance of HPV30. Additionally, enhancement of antibody titers against Newcastle Disease Virus occurred in vaccinated chicks following administration of Phyto V729, further supporting the notion that Phyto V7 has immune-stimulatory properties.

Additional studies should be performed to support the notion that supplementation of phytochemicals to HIV-1 infected patients is beneficial, and to elucidate their mechanism of action. This should be done with larger cohorts of HIV-1 infected individuals and AIDS patients. In the current study only patients with less than 350 CD4+ T-cells per mm³ were recruited. Future studies should examine individuals with significantly higher CD4+ T-cell counts not receiving HAART as well as individuals with very low CD4 T-cell counts receiving or not receiving HAART. Future studies should also include patients receiving just multiple micronutrients supplementation (MMN) and compare them with patients receiving just phytochemicals. If proven the significant added value of phytochemicals over MMN, current recommendations, like the UN requested of inclusion of MMN for treatment of HIV carriers and AIDS patients at any stage of their disease, should be revised to include phytochemicals.