Effect of heat stress on production parameters and immune responses of commercial laying hens.

The present study was conducted to determine the adverse effects of high temperature and humidity not only on live performance and egg quality but also on immune function in commercial laying hens. One hundred eighty 31-wk-old laying hens at peak production were used in this study. Hens were housed in cages (15 cages of 4 birds/cage) in each of 3 environmental chambers and received 1 of 3 treatments. The 3 treatments were control (average temperature and relative humidity), cyclic (daily cyclic temperature and humidity), and heat stress (constant heat and humidity) for 5 wk. Different production and immune parameters were measured. Body weight and feed consumption were significantly reduced in hens in the heat stress group. Egg production, egg weight, shell weight, shell thickness, and specific gravity were significantly inhibited among hens in the heat stress group. Likewise, total white blood cell (WBC) counts and antibody production were significantly inhibited in hens in the heat stress group. In addition, mortality was higher in the heat stress group compared to the cyclic and control groups. Even though T- and B-lymphocyte activities were not significantly affected by any of the treatments, lymphocytes from hens in the heat stress group had the least activity at 1 wk following treatment. These results indicate that heat stress not only adversely affects production performance but also inhibits immune function.

Initiation of humoral immunity. I. The role of cytokines and hormones in the initiation of humoral immunity using T-independent and T-dependent antigens.

The early events during the initiation of immune responses following the injection of T-independent (lipopolysaccharide, LPS) and T-dependent (bovine serum albumin, BSA) antigens were studied in immature male chickens. Specifically, the role of cytokines and hormones in the initiation of humoral immunity against these antigens was investigated. Both interleukin-1 (IL-1) and tumor necrosis factor (TNF-alpha) increased significantly post-LPS but not post-BSA injection. While interleukin-2 (IL-2) significantly decreased post-LPS injection, IL-2 significantly increased post-BSA injection. Furthermore, corticosterone levels significantly increased and tri-iodothyronine (T(3)) levels significantly decreased post-LPS but not post-BSA injection. In this study, the results indicate that although LPS and BSA can induce a humoral antibody response in chickens, they activate different cytokines and neuroendocrine network systems.

Initiation of humoral immunity. II. The effects of T-independent and T-dependent antigens on the distribution of lymphocyte populations.

The effect of injecting T-independent (lipopolysaccharide, LPS) and T-dependent (bovine serum albumin, BSA) antigens on the redistribution of lymphocyte populations in immature male chickens was investigated. In the blood, percentages of total T-cells (CD3+), T-helper cells (CD4+), and T-cytotoxic/suppressor cells (CD8+) significantly decreased post-LPS injection (PLI) but not post-BSA injection (PBI), while percentages of monocytes/thrombocytes (K1+) significantly increased PLI. Interleukin-1 receptor expression on blood lymphocytes increased significantly PLI and PBI. In the spleen, the percentages of total T-cells (CD3+) increased significantly PLI and PBI, macrophage (K1+) percentages increased significantly PLI, while B-cell percentages decreased significantly PLI. These results indicate that following antigen injection, there is a redistribution of peripheral blood lymphocytes (specifically T-lymphocytes) to secondary lymphoid organs and the kinetics and magnitude of the changes can differ according to the type of antigen used.

Effects of photoperiod and melatonin on lymphocyte activities in male broiler chickens.

Understanding the role of the pineal gland in regulating the immune response and the role of photoperiod in influencing pineal gland secretions are becoming increasingly important. The purposes of the present experiments were to investigate the effects of different photoperiod regimens on T- and B-lymphocyte activities in broiler chickens. Next, the influence of different photoperiod regimens on the responsiveness of lymphocytes to melatonin in vitro was examined. The effect of melatonin in vitro on lymphocyte activities was also studied, regardless of the photoperiod received. Finally, the effects of photoperiod on the profiles of different splenocyte cell types were investigated. To study the effect of photoperiod on lymphocyte activities, different photoperiod regimens were used. These were: constant lighting, 23 h light:1 h darkness; intermediate lighting, 12 h light:12 h darkness; and intermittent lighting, 1 h light:3 h darkness. Peripheral blood and splenic lymphocyte activities were tested at 3 and 6 wk of age by performing a mitogen cell-proliferation assay with a polyclonal T-cell mitogen, concanavalin A (Con A), and T-dependent B-cell mitogen, pokeweed mitogen (PWM). To study the effect of photoperiod on the responsiveness of lymphocytes to melatonin in vitro or the effect of melatonin in vitro on lymphocyte activities regardless of photoperiod received, lymphocytes from the chickens that were exposed to the different photoperiod regimens were incubated with mitogen and different concentrations of melatonin. To study the effect of photoperiod on profiles of different cell types, the percentages of splenocyte subpopulations from birds exposed to different photo-periods were determined using flow cytometry with CD4+, CD8+, CD3+, and B-cell markers. The results of these studies indicate that splenic T and B lymphocytes from 6-wk-old chickens grown in intermittent lighting had higher activities than those from chickens grown in constant lighting. Peripheral blood and splenic lymphocytes from chickens raised under constant lighting were more responsive to melatonin in vitro than those from chickens raised under intermittent lighting. This difference in response may be due to lower levels of melatonin in birds receiving constant lighting, making them more sensitive to melatonin in vitro. Melatonin in vitro enhanced the mitogenic response of peripheral blood T lymphocytes from 6-wk-old chickens, splenic T lymphocytes from 3-wk-old chickens, and splenic T and possibly B lymphocytes from 6-wk-old chickens. Finally, intermittent lighting increased the percentages of splenic CD4+, CD8+, and CD3+ cells but not B-cell subpopulations at 6 wk of age, presumably because of increased levels of melatonin in birds receiving intermittent lighting. Our results re-emphasize the importance of melatonin in regulating host immune response; this regulation could be accomplished through exposing broiler chicks to intermittent lighting.

Effect of genetic selection and MHC haplotypes on lymphocyte proliferation and interleukin-2 like activity in chicken lines selected for high and low antibody production against sheep red blood cells.

Chickens from third generation matings of lines of chickens selected for high (HA) and low (LA) antibody production to sheep red blood cells (SRBC) and typed for MHC genotypes B13/13, B13/21, and B21/21 were used in this study. Chickens from both lines carried all the three genotypes B13/13, B13/21, and B21/21. To study T- and B-lymphocytes mitogenic activity, 12-week-old female chickens were injected intravenously with 0.2 ml of 9% SRBC and spleens were collected at 0, 6 h, and 6 day post-antigen injection (pAg). Isolated lymphocytes were incubated with either Concanavalin-A (Con-A) for T-cell activity, or Pokeweed mitogen (PWM) for B-cell activity and thymidine 3H uptakes were measured. To study the Interleukin-2 (IL-2)-like activity in the same lines and genotypes, splenic lymphocytes from 12-week-old chickens were passed through nylon wool columns to enrich the T-cell population. After a 24 h incubation with Con-A, the conditioned media (CM) were collected. The CM were tested for IL-2 like activity by determining whether they altered the proliferation of Con-A stimulated T cells. This proliferation effect was then compared to that of a reference conditioned media (RCM) prepared from K-strain birds and that were used as the standard for the assay. There was no significant difference (p > 0.05) in IL-2 like activity between HA and LA lines, however, the LA was significantly higher than HA (p < 0.05) in T- and B-cell mitogenic activity. The genotype B13/13 had significantly higher (p < 0.05) IL-2 like activity than the B21/21. The genotype B13/13 was also significantly higher (p < 0.05) in T- and B-cell mitogenic activity than the B21/21. At 0 h, pAg T- and B-mitogenic activity was significantly higher (p < 0.05) than 6 h. In summary, our results indicate that although the birds were selected for high antibody production to SRBC, their lymphocyte mitogenic activity was lower than those selected for low antibody production. Hence, humoral and cell-mediated immune responses appear to be under different genetic controls, and that selection for greater humoral response may be at the expense of cellular responses. Our results also suggest differences in IL-2 like activity production between chickens carrying different MHC B-haplotypes, and that genetic control of such activity is possibly linked to the MHC genes.