Sites and trend of erythropoiesis in anemic, normal, and splenectomized newts

Newts, Triturus cristatus carnifex (Laurenti), were anesthetized by submersion in 2% chlorbutol in tap water for 15 min, splenectomized and then rendered totally anemic two months later by treatment with acetylphenylhydrazine (APH) diluted in their tanks (25 mg/liter for 36 h, changing the solution every 12 h). In the 14 weeks following hemolysis, erythron restoration occurred with the same intermittence as it did in whole animals rendered anemic by APH treatment: Beginning the second week the red blood cell count progressively increases for about one month, followed by a period of stasis which lasts about three weeks, then by a new increase, and then by a final period of stasis. Histological examination shows that erythropoietic activity occurs partly in the circulating blood and partly in erythroblasts nestled in the crypts between the muscular trabeculae of the ventricle as well as in the atrial walls. These cells, which are not part of the freely circulating elements in the blood stream, become very abundant in both whole and splenectomized anemic newts but are also present in normal animals. Newts, thus, have three sites for erythropoiesis: the spleen, the blood stream, and the heart. The other components compensate for the elimination of the spleen without determining any lack of, or delay in, erythropoietic response.     

Antifertility effects of immunoglobulins from uterine fluids of semen-immunized rabbits

The effects on fertility of the immunoglobulins, IgG (similar to serum IgG) and secretoy IgA (the predominant IgA in uterine fluid, different from serum IgA) and the fluids from the uteri of semen-immunized rabbits were studied. Female rabbits were immunized either by transvaginal (TV) injections (every 4 or 5 days for 3 weeks) with adjusted semen added to buffered saline containing .5 mg each polyadenylic acid )Poly-A) and polyuridylic acid or a combination of transvaginal injections and systemic injections with rabbit semen mixed with adjuvant (each once weekly for 3 weeks). 2 weeks following the last injection of semen each rabbit received a TV injection followed in 1 week by an instillation of semen and Poly A and U into uterine horns ligated at both ends. 2 weeks following intrauterine (IU) instillation uterine contents were withdrawn. Uteri were reinstilled. Passive hemagglutinating (PHA) and sperm-immobilizing antibody titers were determined on both serum and adjusted uterine fluid (UF) samples. Sperm immobilizin tests and PHA were positive in all serum samples from the contained immunization group, and PHA were positive in 4 out of 7 UF samples of this group. 2 out of 10 and 3 out of 10 samples from the TV group were positive for SI and PHA, respectively, and 4 out of 10 UF samples were PHA positive. Treatment of rabbit semen with UF samples prior to insemination of estrous rabbits resulted in an implantation rate of 2.8% for the combined immunization group and 24.2% for the TV group compared to 79.2% for controls. Some IUF samples without detectable antibodies appeared to inhibit fertility. The implantation rate of sperm treated with immune UF from the TV group absorbed with goat normal serum, anti-rabbit gamma-globulin serum, anti-rabbit-secretory IgA serum, anti-rabbit IgG serum, or sperm, were 3.4%, 88.9%, 42.9%, 64%, and 73. 3% respectively. SIgA and IgG antibodies were seperated by gel filtration. 2 of 4 pooled IgA samples and 3 of 4 pooled IgG samples depressed fertility, with 24.3% and 22.6% implantation and 25.8% 5, and 0% implantation, respectively. To assure that the antifertility effects observed were due to sperm antibodies of each immunoglobin class, SIgA fractions of 2 IUF pools were subjected to anion-exchange chromatography, and both SIgA and IgG fraction were absorbed with goat NS, anti-SIgA, and anti-IgG. IgG fractions absorbed with anti-SIgA or anti-IgG resulted in implantation rates of 0% and 56.8%, respectively, compared to a control of 89.5%. SIgA fractions absorbed with anti-SIgA or anti-IgG resulted in implantation rates of 81.8% and 19.4%. Thus IgG and secretory IgA of the uterine fluids are active in inhibition of fertility.     

Comparative radioprotective efficacy of native and irradiated in vitro immunoglobulins from horse serum

The therapeutic application of native and in vitro exposed (20000 Gy immunoglobulins of horse blood serum increased the survival of irradiated (LD75-95) animals, normalized the quantitative and qualitative status of the small intestine microflora and prevented enterobacteria from penetrating the internal organs. The irradiated preparations were more active than native ones.     

Purification from goat antiserum of immunoglobulins against G6PD from rabbits]

DEAE Affi-Gel Blue (Bio-Rad) provides an efficient and rapid fractionation of human serum proteins by a single chromatographic step. When goat serum is applied to the matrix and chromatography is performed following the procedure utilized for the human serum proteins, the elution pattern changes and the Ig purification is not satisfactory. We achieved a better Ig purification from goat serum by the following improved procedure. We performed first an AS-40 fractionation followed by extensive dialysis in 50 mM Na-citrate pH 5.7. The sample was then loaded onto a P11 column equilibrated in the same buffer. The fraction eluted at Vo contained total IgG and the other serum proteins, except beta-globulins which were eluted with 0.24 M phosphate. Peak 1 concentrated and dialyzed in 20 mM phosphate buffer pH 8 was then applied to a DEAE Affi-Gel Blue column, equilibrated in the same buffer. Two protein peaks were eluted from this column and electrophoretically characterized as: peak 1, containing a pure Ig fraction (70% yield), peak 2 with albumin and other contaminating serum proteins. When goat antiserum is obtained against a specific protein, our technique may be suitably employed to purify polyclonal antibodies for immunoprecipitation studies.     

High-affinity binding of riboflavin and FAD by immunoglobulins from normal human serum

The binding of [3H]FAD and [3H]riboflavin to a pooled, human plasma immunoglobulin fraction was studied. For each flavin, the data fit best a model with two binding sites of high affinity and a class of sites of lower affinity. The dissociation constants estimated for the two high affinity sites were 1.73 nM and 0.078 nM for [3H]FAD and 2.43 nM and 0.068 nM for [3H]riboflavin. The results of studies with a series of possible competitors suggested that the flavin ring system was an important determinant of the binding. Other studies showed that the binding reaction was largely enthalpy-driven. Our findings show that normal human immunoglobulins contain one or more species that bind riboflavin and FAD with very high affinity.     

Babesia argentina: the infectivity and immunogenicity of irradiated blood parasites for splenectomized calves

The intraerythrocytic forms of Babesia argentina were exposed to various doses of γ-radiation and then inoculated intravenously into splenectomized calves to study infectivity and immunogenicity of the irradiated parasites. Commencing with a dose of 8 krads, the proportion of organisms capable of multiplication in the host was progressively reduced until complete inhibition of multiplication was obtained with doses in the range of 50–70 krads. After exposures between 20 and 50 krads, the organisms showed reduced rates of multiplication in the host and produced only mild infections that left the surviving recipient animals strongly immune to reinfection. Parasites, completely inhibited by irradiation, did not protect splenectomized calves against challenge infection and behaved immunologically as killed organisms.