The world has finally settled living with Human immunodeficiency Virus (HIV) with no cure discovered so far. Yong people with HIV infection under HIV management drugs get married and eventually end up being pregnant and bearing babies. The need therefore to investigate the effect of HIV infection and antiretroviral drugs on body chemistry especially the hormones concerned with pregnancy and lactation cannot be over emphasized as this is important to intervene when necessary for the overall benefit for the mother and child. We investigated the effect of anti-retroviral drugs and human immune deficiency virus (HIV) infection on prolactin production and stimulation in HIV-infected pregnant women. A total of 120 subjects participated in the study. Sixty (group 1) of these subjects were HIV seropositive pregnant women who commenced treatment with zidovudine in combination with lamivudine, that is highly anti-retroviral therapy (HAART) at 2 nd trimester. The 2 nd group made up of 60 HIV seronegative pregnant women who received no drug and as control to the study. The blood samples of both groups were collected at the beginning of the second and at the end of the third trimesters. For group 1 (seropositive pregnant women), the blood collection was done just before commencing the HAART treatment. The prolactin level of HIV seropositive pregnant women were significantly (P < 0.05) lower than the HIV seronegative pregnant women at the 2 nd trimester. Also, the prolactin level of HIV seropositive pregnant women at 2 nd trimester was not significantly increased (P> 0.05) compared with third trimester level. The reverse was the case with HIV seronegative pregnant women where prolactin level of 3 rd trimester was significantly increased when compared with 2 nd trimester. HIV infection has prolactin suppressive effect on pregnant women and HAART treatment did not significantly raise prolactin level.