A landmark contribution to poultry science--Immunological function of the bursa of Fabricius.

The article "The bursa of Fabricius and antibody production" by Bruce Glick, Timothy S. Chang, and R. George Jaap first demonstrated the role of the bursa in immune development. Birds, including chickens, possess a peculiar organ, the bursa of Fabricius. The organ was recognized for more than 300 yr before its function was described in 1956. Discovery of the bursa as an essential component of the immune response began by accident. Removal of the bursa, bursectomy, during the rapid growth period diminished the antibody response to Salmonella. A paper describing this exceptional finding was initially rejected by Science and ultimately published in Poultry Science. This revelation triggered sequential events leading to the understanding of the dichotomy of the immune response. Additional work in multiple laboratories over many years revealed fundamental immune mechanisms attributable to the bursa. Understanding those mechanisms advanced agricultural and biomedical science.

Catecholamines modulate chicken immunoglobulin M and immunoglobulin G plaque-forming cells.

Norepinephrine (NE) and epinephrine (E) function as chemical messengers in the central nervous and the endocrine systems of the chicken. The effects of in vivo and in vitro exposure of NE and E on IgM and IgG splenic plaque-forming cell (pfc) formation were determined to the antigen SRBC. Six-week-old Line UNH 105 New Hampshire chickens were injected i.v. with NE (500 micrograms/kg BW) or E (100 micrograms/kg BW) followed by 1 mL of 5% SRBC 30 min later. Five days after antigen injection, IgM and IgG pfc were assayed. Compared with controls, in vivo NE suppressed (P < .05) IgM and IgG pfc formation. In vitro NE treatment of splenic lymphocytes reduced (P < .05) IgM pfc but did not affect IgG pfc numbers. In vivo treatment with E increased (P < .05) IgM pfc whereas in vitro E exposure increased (P < .05) IgM pfc. Immunoglobulin G pfc were suppressed (P < .05) by both in vivo and in vitro E exposure. The presence of surface receptors for NE and E on splenic lymphocytes was determined using in vitro incubation with antagonists to alpha and beta receptors. These data suggest that there are alpha and beta receptor sites on lymphocytes for NE and E, and that these catecholamines have a regulatory role in plaque cell proliferation.

Profile of chicken macrophage functions after exposure to catecholamines in vitro.

The effects of catecholamines (CA) on various chicken macrophage functions were examined. Macrophage monolayers were exposed to .01, .1, .25, 1, 2, and 5 micrograms/mL of dopamine (DA), norepinephrine (NE) and epinephrine (E) for 1 hr. All CA were toxic for macrophages at 1-5 micrograms dose range resulting in 25-50% cell death. All CA at the .1 and .25 micrograms/mL level increased E. coli and sheep red blood cells (SRBC) phagocytosis by macrophages. The percentage of Fc-receptor positive macrophages increased after CA exposure. Prolonged exposure of macrophages (3 hr) reduced SRBC phagocytosis by DA-treated but not in NE- and E-treated macrophages. However, after 1 hr exposure and 3 hr recovery period, CA-induced changes were reversed in all but DA-treated cultures. Apomorphine and metoclopromide blocked DA whereas propranolol blocked NE and E effects suggesting specificity of the observed effects via catecholaminergic receptors on chicken macrophages. Dopamine and NE (.25 micrograms/mL) did not affect but E exposure enhanced LPS-induced tumoricidal factor production. These findings suggest that CA modulate chicken macrophage effector functions.

Continuous administration of dopamine alters cellular immunity in chickens.

Dopamine administered continuously through osmotic pumps altered the PHA wattle response and in vitro leukocyte capillary tube migration in UNH 105 chickens. The PHA wattle response was suppressed significantly by 48 hr exposure to dopamine at a dose of 1 microgram/hr. Administration of 10 micrograms/hr dopamine for 48 hr enhanced significantly in vitro leukocyte migration.

Biogenic amines regulate avian immunity.

Alzet mini-osmotic pumps were implanted subcutaneously to administer norepinephrine (NE) and epinephrine (E) to 6-wk-old line UNH 105 chickens. Dose-time studies showed the most effective NE and E dose and exposure time on two chicken cellular immune responses: the phytohemagglutinin (PHA) wattle response and leukocyte migration. Administration of 1 microgram/h NE for 72 h suppressed significantly the wattle stimulation index [2.48 +/- .3 (SE)] compared to that of saline controls (4.1 +/- .3) but enhanced mean leukocyte migration (7.7 +/- .3 versus 4.9 +/- .3). Epinephrine at 1 microgram/h for 72 h significantly suppressed the wattle index (1.8 +/- .2) compared to that of controls (2.8 +/- .3) but E at 1 microgram/h for 48 h enhanced leukocyte migration (9.5 +/- .2 versus 6.4 +/- .2). Continuous administration of NE and E at physiological levels alters cell-mediated immunity and appears to have an immune regulatory role in the chicken.

Modulation of chicken plaque-forming cells by serotonin and dopamine.

Serotonin (5-hydroxytryptamine, 5-HT) and dopamine (DA) are endogenous components of the central nervous and endocrine systems of the chicken. To determine the effects of these monoamines on antibody-mediated immunity. New Hampshire chickens of Line UNH 105 were injected intravenously with 5-HT (100 micrograms/kg of body weight) and DA (1 mg/kg of body weight). One milliliter of a 5% SRBC suspension was injected intravenously 30 min later. Both IgM and IgG splenic plaque-forming cells were assayed 5 days after antigen injection. For in vitro studies, spleen lymphocytes from SRBC-primed chicks were incubated with DA and 5-HT followed by quantitation of IgM and IgG plaque-forming cells. The in vivo incubation of splenic lymphocytes with specific antagonists was used to ascertain the presence of monoamine receptors on lymphocytes. The 5-HT significantly enhanced IgM plaque-forming cells compared with controls following in vivo [550 +/- 85 (SE) cells/10(6) splenic lymphocytes versus 359 +/- 44] but not in vitro exposure. The IgG plaque-forming cells were not affected by 5-HT. The DA significantly suppressed IgM plaque-forming cells responses following in vivo (284 +/- 46 versus 499 +/- 66) and in vitro (254 +/- 57 versus 451 +/- 51) exposure. Significant suppression of IgG plaque-forming cells was found in vivo (287 +/- 40 versus 462 +/- 75) and in vitro (153 +/- 36 versus 371 +/- 81) following treatment. Specific DA antagonists, apomorphine and metoclopramide, did not alleviate the in vitro suppressive effect of DA.

Monoamines alter in vitro migration of chicken leukocytes.

The effect of the biogenic amines serotonin (5-HT), dopamine (DA), and norepinephrine (NE) on peripheral blood leukocyte (PBL) migration was studied in two populations derived from line UNH 105 New Hampshire chickens. Maximum migration from capillary tube migration chambers was achieved in 1 hr. An age effect in both populations was indicated by significantly larger migration areas found in leukocytes from 7-week-old chickens compared to those of 4-week-old chicks. Thirty min after intravenous monoamine injection, line UNH 105 PBL migration was unaffected by exogenous monoamines. In the second population, B24/B24 chicks, NE enhanced migration at 4 weeks of age but DA suppressed migration at 7 weeks of age. In vitro exposure of PBL to the biogenic amines also affected leukocyte migration. Migration was augmented by 100 ng 5-HT but suppressed by 1 microgram 5-HT in UNH 105 chicks. Furthermore, DA suppressed PBL migration and NE enhanced migration in the same population. PBL from B24/B24 chicks were not affected by in vitro exposure to 5-HT, however, DA enhanced migration whereas NE suppressed migration. Specific antagonists for 5-HT, DA, and NE blocked the effects of each monoamine suggesting that receptors are present on chicken leukocytes. These receptors mediate action of the monoamines on leukocyte migration activity.

Continuous administration of 5-hydroxytryptamine alters cellular immunity in chickens.

1. 5-Hydroxytryptamine administered continuously through osmotic pumps altered the PHA wattle response and in vitro leukocyte capillary tube migration in UNH 105 chickens. 2. Administration of 100 ng/hr 5-hydroxytryptamine for 48 hr significantly suppressed the PHA wattle response. 3. In vitro leukocyte migration was significantly enhanced by 72 hr exposure to 5-hydroxytryptamine at a dose of 100 ng/hr.

Ontogeny of antibody response to Brucella abortus in turkeys.

No differences were detected between large White Nicholas and small White Hybrid turkeys in antibody (AB) titers in birds vaccinated with Brucella abortus. Maximal AB titers occurred by 6 weeks of age and remained high throughout life. B. abortus elicits an AB response when injected at day of hatch while bursectomy at day of hatch and 2 days of age significantly depressed this response. There was an apparent immunological maturation process occurring to B. abortus in the bursa of Fabricius which contained for approximately 6 weeks.

Effects of Bordetella avium on lymphoid tissue monoamine concentrations in turkey poults.

Six hours post-hatch, large white turkey poults were inoculated intranasally with 5 x 10(7) cells of the W isolate of Bordetella avium. Three hours after inoculation and subsequently on days 7, 10, 14, 21, and 28 postinoculation, poults from infected and control groups were killed by cervical dislocation. Thymus, spleen, and bursa of Fabricius were removed, weighed, and frozen until assayed for norepinephrine (NE), dopamine (DA), and serotonin (5HT). B. avium infection caused a reduced concentration of NE, DA, and 5HT in the spleen, thymus, and bursa of Fabricius. The reduced concentrations of these monoamines in lymphoid tissues of diseased poults may be a normal response during the course of a disease or during the mounting of an immune response.

Brain monoamine concentrations in turkey poults infected with Bordetella avium.

Six hours post-hatch, large white turkey poults were inoculated intranasally with 5 X 10(7) organisms of the "W" isolate of Bordetella avium. Three hours after inoculation and subsequently on days 7, 10, 14, 21, and 28 postinoculation, 30 infected and 30 uninfected control poults were injected intramuscularly with alpha methylparatyrosine (AMPT, 250 mg/kg), and an additional 30 of each group received a saline vehicle. Three hours after AMPT injection, whole brains from treated and untreated poults from both infected and control groups were removed, weighed, and frozen until assayed for norepinephrine (NE), dopamine (DA), and serotonin (5-HT). B. avium-infected poults had a significant reduction in steady-state NE and Da and a greater depletion of NE and DA after AMPT treatment compared with control poults. Infected poults had significantly reduced whole brain 5-HT concentrations, which persisted through 21 days postinoculation. Altered brain NE, DA, and 5-HT concentrations suggest that the B. avium-infected poults may be less able to cope with additional stressors.

Monoamines suppress the phytohemagglutinin wattle response in chickens.

The influence of the biogenic amines serotonin (5-HT), dopamine (DA) and norepinephrine (NE) on the phytohemagglutinin (PHA) wattle response was studied in three populations derived from line UNH 105 New Hampshire chickens. At 3 or 6 weeks of age, chicks were injected intravenously with either 5-HT, DA, NE, or a saline control. Thirty minutes after monoamine injection, 100 ug PHA-P was injected in the wattle and the thickness increase was calculated 24 hr later. Analysis of variance showed significant monoamine effects compared to controls but no difference in response among the three populations. 5-HT and NE each depressed significantly the wattle response in 3 and 6 week old chicks. DA reduced the response in both ages also but the significantly greater reduction in 6 than in 3 week old chicks indicated an age effect. These results suggest a regulatory role for monoamines in the PHA wattle response.

Alcaligenes faecalis infection in turkeys: effects on serum corticosterone and serum chemistry.

The serum levels of corticosterone, total calcium, inorganic phosphorus, protein, sodium, and potassium were determined for turkey poults from 1 to 42 days of age. Corticosterone levels were higher at days 10, 14, 21, and 28 in Alcaligenes faecalis-infected turkeys than in uninfected poults. The corticosterone responses in this study indicate that A. faecalis infection stimulates adrenal cortical function even in the very young poult, which ordinarily does not show adrenal cortical responsiveness to various environmental and physical stressors. No differences were observed between infected and uninfected turkey poults in other serum chemistries.

In vitro cellular migration of leukocytes from turkey poults infected with Alcaligenes faecalis.

The ability of peripheral blood leukocytes from young turkey poults to migrate in vitro was investigated. Migration from capillary tubes was relatively rapid and was usually complete in 2 hours. Leukocyte migration was significantly enhanced in Alcaligenes faecalis-infected turkeys compared with uninfected controls at 1, 2, 5, and 6 weeks of age.

Body temperature response of turkey poults infected with Alcaligenes faecalis.

Experiments were conducted to investigate the colonic temperature (TC) response to Alcaligenes faecalis infection in turkeys. One-day-old poults were inoculated intranasally with 0.05 ml of a 24 hour brain heart infusion (BHI) broth culture with a bacterial concentration of 1.0 x 10(9) cells/ml. Within 2 hours after inoculation TC was depressed significantly in challenged poults. This depression was significant through day 14 in experiment 1 and day 6 in experiment 2. Further studies with sterile BHI broth, BHI broth culture supernatant and phenol killed bacteria indicated that TC depression occurred only with the killed bacteria. This TC depression with killed bacteria lasted 3 days. Foot pad temperature (TF), an indicator of peripheral vasomotor tone, was depressed through day 9 in A. faecalis-infected poults. The reduced TC and TF of infected poults is highly suggestive of a reaction to an endotoxin in young birds. The mechanisms whereby A. faecalis allows poults to become more susceptible to other infective agents may reside in physiological changes leading to reduced body temperature.

Immediate hypersensitivity (anaphylaxis) response in uninfected and Alcaligenes faecalis-infected turkey poults.

The immediate hypersensitivity (anaphylaxis) response in uninfected and Alcaligenes faecalis-infected turkey poults was studied. The optimum challenge dose of bovine serum albumin (BSA) to elicit an anaphylactic response was 40 mg/kg body weight, and the optimum poult age was 4 to 5 weeks. The response latency time was 7 days. The anaphylactic response was specific for the sensitizing antigen used. No differences were observed between A. faecalis-infected and uninfected poults in the immediate hypersensitivity response to BSA.

Anti-sperm antibodies and fertility of turkey hens.

Anti-sperm antibody titers increase with time in serum of turkey hens following a standard production schedule of artificial insemination (AI). In hens receiving intravenous (IV) or intraperitoneal (IP) additional AI, serum anti-sperm antibody levels increase more rapidly after a lag phase. A single injury to the oviduct also resulted in increased anti-sperm antibodies similar to IV and IP groups. This is a new observation that a single injury increased antibody titers to spermatozoa equal in IV and/or IP injections. A negative correlation between serum anti-sperm antibody titers for IV, IP and injury to oviduct and fertility of these groups was observed. Hens of IV and injury to oviduct groups with high levels of anti-sperm antibodies in the last 2 weeks of production had significantly lower fertility than hens with low levels of antibodies and control hens.

Pathogenicity of various isolates of Alcaligenes faecalis for broilers.

Day-old broilers or specific-pathogen-free chickens were inoculated intranasally with approximately 1 X 10(8) organisms of eight different field isolates of Alcaligenes faecalis. Major differences in the pathogenicity of isolates and their ability to colonize the trachea were found. Only two isolates (Wilson and Lockamy) produced mild clinical signs of respiratory disease ("snicking," dyspnea). The same two also colonized the respiratory tract, especially the trachea, in large numbers; they persisted for 31 days. Of the remaining six isolates, five were also able to colonize the respiratory tract but did so to a lesser degree and less persistently, without causing clinical signs. Only one isolate (CS) was incapable of becoming established in the respiratory tract of chicks after intranasal inoculation.