Hepatitis B virus (HBV) is a significant public health concern, particularly in low-income countries. The host immune response plays an essential role in hepatitis B virus outcome, mainly orchestrated by cytokines and immune modulators molecules such as vitamin D. This study aimed to determine the levels of cytokines (IL-10), interferon-gamma (IFN-gamma), interferon-lambada (IFN-lambada), granzyme B, and vitamin D in Khartoum, Sudan, and to assess the association between them and variable HBV viral load as well as liver enzyme levels. A total of 174 participants were enrolled in the trial, with 100 HBV-infected patients (who were HIV and HCV negative) and 74 healthy volunteers. Patients’ HBV viral load, vitamin D levels, liver enzymes, and circulatory cytokines were measured. This study revealed a remarkable decrease in vitamin D levels in HBV infected patients with an elevation in levels of liver biomarkers. The level of IL-10 was significantly higher in patients than in apparently healthy controls (p-value <0.001). There was a positive correlation between IFN-ϒ, Granzyme B, and viral load. According to the findings of this study, there is a marked hypovitaminosis D among hepatitis B infected patients with a significant increase in IL-10, which may implicate the persistence of HBV infection. Moreover, HBV DNA levels were significantly associated with IFN- g and granzyme B levels.