Polychlorinated biphenyls (PCBs) are persistent organic pollutants (POPs) causing adverse effects on both humans and animals such as endocrine disruption, neurotoxicity and immunomodulation. Immune system is an important target organ for many environmental contaminants. Macrophages represent an important component of the immune system of humans and animals; in fact, they are responsible for triggering innate immune response and host defence. Although it is well known that different environmental pollutants may affect immune function, only few studies have been carried out to date about the effects of PCBs in macrophages; while, at our knowledge, no study investigated the effects of these contaminants on porcine macrophages. The aim of the current study was to evaluate the effects of three non dioxin like PCB congeners (PCB 138, PCB 153 and PCB 180) alone and differently combined, on porcine macrophages cell line (3D4/31). The results of the current study showed that ndl-PCBs reduced significantly cell viability only at the highest tested concentrations (50 μM); such effect was not linked to apoptosis induction or cell cycle arrest. The contemporary presence of more than one contaminant (differently combined) did not induce any enhancement of effects on 3D4/31 cell line. These data highlighted the need to continue the evaluation of toxic properties of ndl-PCBs, which represent less studied PBCs, in particular in term of risk assessment.