The pig is a useful model for the heterogeneity of the mammalian immune system and has also recently received attention as a possible source of organs for human transplantation. Here we report a detailed analysis of porcine lymphocyte phenotypes. Peripheral blood αβ T cells consisted of four subsets (CD4+8−, CD4+8lo, CD4−8lo and CD4−8hi) and γδ T cells of three (CD2−4−8−, CD2+4−8lo and CD2+4−8−). There were, in addition, a large proportion of non‐T‐non‐B lymphocytes with CD2+3−4−8lo surface immunoglobulin‐negative phenotype containing natural killer (NK) activity. A striking observation was the relatively low frequency of αβ T cells in the blood of young pigs. Similar phenotypes were also identified in the cells from peripheral lymphoid tissues, though the proportions of the γδ T cells and the non‐T‐non‐B lymphocytes in the lymph nodes and tonsil were much lower and the majority of the γδ T cells in the lymphoid tissues bore CD2 and/or CD8. In thymus, the small thymocytes were predominantly CD3−4+8+while the mature large thymocytes displayed phenotypes similar to those of peripheral T cells. Thus this work has directly defined porcine αβ and γδ T cells, demonstrated the T‐cell nature of the unique CD4+8+ subset of peripheral lymphocytes, revealed the high heterogeneity of the CD8+ cells, and established the phenotype of NK cells. The functional properties of these defined porcine lymphocyte subsets can now be experimentally determined in health and disease.

Although porcine lymphocytes have been classified into numerous subpopulations in postnatal animals, little is known about the ontogeny of these complex cell subsets. Using double- and triple-colour flow cytometry (FCM), we investigated the surface phenotype of fetal lymphoid cells in the thymus, cord blood, spleen and mesenteric lymph nodes at different stages of gestation. It was found that the major lymphocyte subpopulations started to appear at the beginning of the second third of the gestation period, with B cells being the earliest lymphocyte subpopulation to appear in the periphery. The T-cell receptor (TCR) gamma delta+ cells were the earliest detectable T-cell subset, developing first in the thymus and subsequently arriving in the periphery. Later in ontogeny, however, the number of TCRalpha beta+ lymphocytes rapidly increased, becoming the predominant T cells both in the thymus and in the periphery. Cells with the phenotype of adult natural killer cells were also identified in pig fetuses, though their nature and functional roles remain to be investigated. In addition, CD2 was expressed on most B cells whilst very few CD4+ TCRalpha beta+ cells or CD2+ TCRgamma delta+ cells expressed CD8, suggesting that the expression of CD2 and CD8 may reflect the functional status of the cells in postnatal animals. Taken together, this study has provided a systematic analysis of fetal porcine lymphocyte subpopulations and may provide the base for studies to establish the physiological roles of these lymphocyte subsets.

Infection of cells with picornaviruses can lead to a block in protein secretion. For poliovirus this is achieved by the 3A protein, and the consequent reduction in secretion of proinflammatory cytokines and surface expression of major histocompatibility complex class I proteins may inhibit host immune responses in vivo. Foot-and-mouth disease virus (FMDV), another picornavirus, can cause persistent infection of ruminants, suggesting it too may inhibit immune responses. Endoplasmic reticulum (ER)-to-Golgi apparatus transport of proteins is blocked by the FMDV 2BC protein. The observation that 2BC is processed to 2B and 2C during infection and that individual 2B and 2C proteins are unable to block secretion stimulated us to study the effects of 2BC processing on the secretory pathway. Even though 2BC was processed rapidly to 2B and 2C, protein transport to the plasma membrane was still blocked in FMDV-infected cells. The block could be reconstituted by coexpression of 2B and 2C, showing that processing of 2BC did not compromise the ability of FMDV to slow secretion. Under these conditions, 2C was located to the Golgi apparatus, and the block in transport also occurred in the Golgi apparatus. Interestingly, the block in transport could be redirected to the ER when 2B was coexpressed with a 2C protein fused to an ER retention element. Thus, for FMDV a block in secretion is dependent on both 2B and 2C, with the latter determining the site of the block.