Guinea Pig Red Blood Cells Research Articles

B and T cells in a cat with thymic lymphosarcoma.

The mixed antiglobulin reaction and the formation of nonimmune rosettes with guinea-pig red blood cells (RBC) distinguished feline B and T cells, respectively. In a cat with thymic lymphosarcoma, the cells reacting in these tests formed separate, nonoverlapping populations. The malignant cells were large lymphoblasts replacing the normal thymus and infiltrating local lymph nodes, where they localized only in the paracortical, i.e., thymus-dependent areas. Cells from the nodes could therefore be identified as malignant or normal by their size. The mixed antiglobulin reaction showed that the malignant cells did not carry the surface Ig characteristic of B cells, whereas these malignant cells formed nonimmune rosettes with guinea-pig RBC. Among lymph node cells, most surviving normal and small lymphocytes, from outside the thymus-dependent areas, reacted as B cells. The morphologic evidence therefore corroborated the test results, which indicated that the formation of rosettes with guinea-pig RBC seems a reliable means for the demonstration of T cells in the cat.

Chemical structure of glycolipid of guinea pig red blood cell membrane.

A hexosamine-containing glycolipid, comprising about 6.8% of the total lipids, was extracted and purified from the guinea pig red cell membrane. The carbohydrate moiety consisted of glucose, galactose, and N-acetylgalactosamine in equimolar proportions. Analysis of the sugar linkages by methylation of this glycolipid indicated that the C-4 positions of glucose and galactose were substituted by another sugar and N-acetylgalactosamine resided on the non-reducing terminal. The configuration of the glycosidic linkages was established by optical rotation measurements, PMR spectra and specific enzymatic hydrolysis. The structure of this glycolipid was shown to be N-acetylgalactosaminyl β(1→4)-galactosyl-β(1→4)-glucosyl-(1→1)-ceramide. The major fatty acid was C24:0, and the long chain base was mostly 4-sphingenine.

Mutant of Polyoma Virus with Impaired Adsorption to BHK Cells

A mutant of polyoma virus PY235 has an impaired adsorption to guinea pig red blood cells and BHK-21 hamster cells. Adsorption to 3T3 mouse cells is much less inhibited. These altered adsorption properties are responsible for the apparent inability of PY235 to cause cell transformation or hemagglutination.

Chagasic cardiopathy. Demonstration of a serum gamma globulin factor which reacts with endocardium and vascular structures.

Twenty-four out of 25 patients with Chagas' heart disease have circulating immunoglobulins which react by indirect immunofluorescence technique with endocardium, interstitium and blood vessels of the heart. With skeletal muscle the reaction was observed in interstitium and vascular structures, but with other organs it was limited to vascular structures. This endocardial-vascular-interstitial factor (EVI) fixed complement. Some evidence indicated that this reaction could be obtained using the serum and tissues from the same patient: for instance, in one positive case a right atrium biopsy was performed. When this substrate was used for indirect immunofluorescence, employing the patient's own serum, positive results were obtained. Specificity is not related to AB blood group systems, or to Forssman or Wassermann antigens. The reacting factor was effectively absorbed from sera with organ homogenates, and with guinea pig red blood cells although it was independent of heterophil antibodies. In almost all cases studied, the EVI factor of the serum, when absorbed with epimastigotes of T. cruzi , results in a negative reaction, suggesting that the genesis of the reacting gamma globulin is related to antigens of T. cruzi. The EVI factor was also observed in 19 of 47 asymptomatic controls from an endemic area with positive serology for T. cruzi and in 3 of 27 with negative serology. These 3 cases had anti- T. cruzi antibodies in titers just below those considered of clinical value. The EVI factor was not observed in 119 normal individuals and 286 patients with selected cardiovascular diseases or another pathology from a nonendemic area. These findings and those mentioned above were statistically significant ( P < 0.001). These results indicate the possibility of a more accurate diagnosis of chagasic myocardiopathy based on the study of the EVI factor, because in an individual case the diagnosis of chronic chagasic cardiopathy can be considered with a low probability in the absence of this factor.

L-Pyrrolidone Carboxylic Acid Content in Mammalian Epidermis and Other Tissues

The total pyrrolidone carboxylic acid (PCA) and free amino acid concentration was determined in epidermis of five mammalian species, as well as in other tissues and body fluids of albino guinea pigs. PCA was found to be a major soluble nitrogen compound in epidermis of all species studied with concentrations of 186, 50, 31, 21, and 19 μmoles/gm wet wt in guinea pig, human, dog, rat, and mouse, respectively. Much lower levels of PCA (2-4 μmoIe/gm wet wt) were present in guinea-pig kidney, liver, pancreas, spleen, intestine, and brain. Guinea-pig red blood cells contained about 0.8 μmole PCA/gm and plasma about 0.3 μmole/ml. Optical rotatory dispersion studies on purified PCA isolated from albino guinea-pig epidermis showed it to be the L isomer. The possible origins of epidermal PCA are discussed.

Capsid components of the parvovirus H-1.

SummaryDisruption of purified H-1 virions with sodium dodecyl sulfate (SDS) and mercaptoethanol (ME) at 100° and pH 7.2 results in the appearance of one major and two minor migrating components on polyacrylamide gels. Molecular weights of 92, 000 (A), 72,000 (B), and 56,000 (C) were determined for the three moieties by coelectrophoresis of the dissociated virus with reference proteins of known molecular weight. Electrophoretic separation of 14C- or 3H-labeled virus polypeptides showed the relative amount of each radioactive component to be 15% (A), 75% (B) and 10% (C) of the total viral protein. Elution of the components from the gels, followed by hemagglutination assay against guinea pig red blood cells, indicated that component B was the antigenic component and structural protein. Amino acid analyses of H-1 virions revealed an overall acidic nature of the protein (acidic/basic amino acids = 1.75).The authors thank Miss Barbara Madden for technical assistance and Mr. Chad Shepis, University of Vermont, f...

Hemagglutination Inhibition with Arboviruses: Relationship Between Titers and Source of Erythrocytes

Antigens for Grand Arbaud, Hazara, and California arboviruses were able to agglutinate goose and either dog, hamster and guinea pig, or hamster red blood cells (RBC) to the same titer at the same pH; in hemagglutination-inhibition (HI) tests, titers for homologous and related sera were the same with these different types of RBC or occasionally one dilution higher with the mammalian cells. Antigens for St. Louis encephalitis and Eastern equine encephalitis viruses required use of lower antigen dilutions with human, guinea pig, and hamster RBC than with goose RBC. The results of comparative HI testing with these latter antigens and types of RBC indicate that HI titer is not directly related to the antigen dilution used with different types of RBC.

Avian Reoviruses. I. Isolation and Serological Characterization

SUMMARY Three virus isolates, viruses 24, 25, and 59, having characteristics similar to reoviruses, were isolated from the chicks showing severe cloacal pasting. The isolation was possible on the chorioallantoic membrane (CAM) of the embryonating chicken eggs, where they multiplied to a high concentration and produced severe edema, enormous thickening and extensive necrosis of the CAM. The embryos of the infected eggs were stunted, and their livers showed necrotic foci. Viruses 24 and 25 were found to be serologically related to each other, whereas virus 59 was unrelated to virus 24 and 25. The viruses had no hemagglutinins for sheep, rabbit, guinea pig, rat, or chicken red blood cells.

Nariva virus: further studies, with particular reference to its hemadsorption and hemagglutinating properties.

Nariva virus induced syncytium formation in BHK-21 and Vero cultures. Such infected cultures showed positive hemadsorption for guinea pig red blood cells (RBC). A hemagglutinin for guinea pig RBC was extracted from infected Vero cells by sonication. The hemagglutinin did not act on 1-day chick or goose RBC, and it showed an enhancement in titer upon treatment with Tween-ether. The hemagglutination reaction was highly temperature-dependent, with optimal titers being obtained at 4° and no activity manifested at 37°. Serological studies by hemagglutination-inhibition and complement-fixation indicated no antigenic relation to the better known myxoviruses or to a number of other viruses. The nucleic acid component of Nariva virus is probably RNA.

Peroxide formation by mycoplasmas which infect man.

SummaryThe hemolytic activity of 8 human and 3 animal species of mycoplasmas was studied. With the exception of T-strains of human origin, each of the mycoplasmas tested hemolyzed guinea pig red blood cells.

Characterization of Mycoplasma pulmonis variants isolated from rabbits. I. Identification and properties of isolates.

Mycoplasma showing at least two colony types were isolated from the nares and oropharynx of New Zealand white rabbits. Two strains were purified by single-colony passages and characterized. Morphology by phase-contrast and electron microscopy was typical of Mycoplasmataceae. Both grew anaerobically as well as aerobically, caused hemolysis of guinea pig, sheep, and horse red blood cells, and fermented glucose. These characteristics are shared by members of the species M. pulmonis, commonly isolated from the respiratory tracts of laboratory rats and mice. By use of the growth-inhibition test and agar-gel double-diffusion tests, the two strains were found to be serologically related to each other and to M. pulmonis ATCC 14267 but not to other representative Mycoplasma species from man and animals.

マウス気管支より分離したマイコプラスマの性状 (第1報)

Since mA strain, one of the isolates from the bronchial tract of normal mice, grows readily in the PPLO broth and produces hemolysin, the biological studies on this strain will help a foundamental understanding of M. pneumoniae on which our experiments are being carried. At present, we described about the isolation of PPLO from the mice, and the growth characteristics and some biological properties with mA strain. The results are summarized as follows.1) PPLO was isolated from 16 cases out of 31 normal 10-month-old mice and none out of ten 3-week-old mice. All of the isolates were recovered from the bronchial tract and none from the heart blood. The titer of the PPLO in 10% suspension of the bronchial tract from which the PPLO was successfully isolated was between 104 and 105 per ml.2) The growth of mA strain in Chanock's PPLO broth was excellent. The maximum titer reached 108 per ml by 2nd day after inoculation, then the titer decreased rather rapidly and the viablity was lost completely by 8th day.3) Carbohydrate utilization pattern of mA strain was the same as that of M. pulmonis.4) The mA strain did not grow in an anaerobic condition and produced α hemolysin for the guinea pig red blood cells.

Precocious anti-diphtheria response induced by RNA immuno-carrier.

IN previous papers an increase of the RNA content in the serum of rabbits immunized with horse serum albumin and rat or guinea-pig red blood cells was observed; the greatest increase was in the gamma-globulin fraction. The RNA extracted from immune rabbit sera (RNA Immuno-Carrier or RNA-I-C) was able to induce in normal rabbits a precocious antibody production against the same antigens used for immunizing the animals' source of the RNA1,2.

MALIGNANT TRANSFORMATION BY POLYOMA VIRUS.

Since the pioneering work of Rous. most research on tumor viruses has been carried out by oncologists rather than by virologists. However, with the recent rapid development of new knowledge concerning the basic mecha­ nisms involved in cytolytic infections with animal viruses, the attention of virologists has been turned to the same questions in the transformation of normal animal cells to malignant cells by the action of tumor viruses. Yet. even now, the amount of such research is limited chiefly because of a lack of available tumor virus-cell systems which lend themselves to modern molecu­ lar biological techniques. The polyoma virus first isolated and studied by Stewart et al. (1) has been investigated by a number of virologists for two reasons. First, it is a more interesting virus since it has the potential of producing a large variety of tumor types in its natural animal host [Stewart et al. (2)]; and, secondly because it can be studied quantitatively by modern virological methods. Although capable of causing malignant transformation in both in vivo and in vitro systems it nevertheless produces a cytolytic infection under certain conditions, and, therefore, can be quantitated in tissue cultures using the plaque technique [Dulbecco & Freeman (3)]. Furthermore, being able to agglutinate guinea pig red blood cells, it can be quantitated by hemagglutina­ tion and through the hemagglutination inhibition (HI)! test can be specifi­ cally identified with ease [Eddy et al. (4)). Since its natural host is the mouse, the availability of many susceptible inbred lines of laboratory mice make it possible to relate in vitro tissue culture phenomena to tumor production in the intact whole animal. This paper will review and evaluate the more important recent research contributions concerning mechanisms of polyoma virus transformation, the relationships between virus and the cell after transformation, and some of the virus-specific properties of the transformed cells. These items will first be considered for the �n vivo transformation and then for that occurring in in vitro systems.

Studies on the Role of Leukocytes in Infection with Influenza Virus

Summary A comparison was made of the associative reactions of PR8 strain of influenza virus with guinea pig red and white blood cells. Leukocytes adsorb virus more efficiently than do erythrocytes, but only a small proportion of the virus taken up by leukocytes was eluted. WBC-associated virus was less available to antibody than was RBC-associated virus, as judged by adsorption of antibody from solution and by neutralization of virus. The leukocytes of peritoneal exudates appeared to contain a higher content of virus than blood obtained from infected mice. Phagocytosis of virus-fluorescein labeled antibody complexes by leukocytes was clearly demonstrated but tissue culture cells showed only surface localization of antigen. The appearance of both types of cells treated with the hemagglutinin subunits of influenza virus obtained by extraction with ether was indistinguishable from cells treated with virus. The results of the studies with fluorescein labeled antibodies were obtained by the application of new techniques developed in the course of this investigation. The significance of the findings is discussed and the hypothesis is presented that leukocytes may serve to transport virus to extrapulmonary sites in infections of man and animals.

Utilization of sheep red blood cells in the haemadsorption test for myxoviruses.

THE hæmadsorption test1,2 has been found to be a simple and useful means of detecting foci of infection by myxoviruses in tissue culture monolayers. The same response may be observed in cell cultures whether these viruses are non-cytopathic3 or cytopathic. Briefly, the technique consists of decanting the growth fluid and covering the monolayer with a 0.4 per cent saline suspension of guinea pig or chicken red blood cells and allowing hæmadsorption to proceed for 10 min. at 4° C. After two saline washes, the areas of hæmadsorption are examined under low magnification.

A heat-stable hemagglutinin in mouse tissue extracts.

SummaryExtracts of normal and neoplastic mouse tissues were found to agglutinate mouse and guinea pig red blood cells. Characteristics of the heat-stable substance(s) responsible for this phenomenon are discussed and it is proposed that the hemagglutinating agent(s) is not a tumor virus.

The electrophoretic properties of red blood cells after reaction with influenza virus hemagglutinin

1. 1. The mobilities of guinea pig red blood cells subjected to the action of PR8 influenza type A virus hemagglutinin have been measured at several values of pH and ionic strength ( pH 7.9−5.7, Γ 2 0.172−0.0043 ). 2. 2. The negative surface charge density of “treated” cells is usually less than that of control cells under similar conditions; at pH 7 and physiological ionic strength the difference amounts to about 1000 e.s.u. or 2 × 10 12 electrons/sq. cm. 3. 3. The reciprocal surface charge density of treated cells varies linearly with reciprocal ionic strength; the slope K is almost the same as for control cells. 4. 4. The charge density of treated cells becomes sensitive to pH at low ionic strengths; the resulting curves are quite similar to those of control cells under the same conditions, but with a constant displacement in the direction of a smaller net negative charge density. 5. 5. The changes associated with receptor decomposition are attributed to the formation of free cationic groups comparable to those formed by the action of RDE upon Tamm and Horsfall's urinary inhibitor. 6. 6. It is suggested on the basis of rough calculations that the red blood cell receptor for influenza A virus consists of a single molecule comparable to the urinary inhibitor. 7. 7. Since the supplementary positive charge on the red blood cell produced by virus action varies with ionic strength in the same manner as the net negative charge of the untreated cell, it is assumed to be attached to the same labile elements of the cell membrane and to participate in the same structural rearrangement brought about by decreased ionic strength.

The electrophoretic properties of red blood cells: The effect of changing pH and ionic strength

1. 1. The electrophoretic mobility of washed guinea pig red blood cells has been measured in solutions of ionic strength 0.172-0.0043 and pH 7.9–5.7. 2. 2. These and other available data lead to the conclusion that the surface charge density of red blood cells decreases at low ionic strengths. 3. 3. At physiological ionic strength, the mobility is nearly independent of pH. As the ionic strength is lowered, the variation with pH becomes more marked, until at ionic strength 0.007 the results obtained after the initial slow adjustment is completed ( ca. 8 min.) resemble the titration curve of a protein. 4. 4. The results are attributed tentatively to changes in the character and position of ionizable groups in the cell membrane, rather than to anion adsorption.

LOCAL ORGAN HYPERSENSITIVENESS

1. Rabbit eyes sensitized with guinea pig red blood cells or fresh egg white respond with an inflammatory reaction following the intravenous injection of the homologous antigen, but not the heterologous. 2. Two-tenths of 1.0 cc. of a multiple antigen containing ten separate ingredients, or in other words, 0.02 cc. of each foreign protein, when introduced into the rabbit's anterior chamber, is sufficient to produce an altered ocular reactivity such that when 1 cc. of one of the ten antigens is introduced intravenously the eye shows hyperemia of the iris and conjunctiva with more or less edema and lacrimation during the next 24 hours. 3. For as long as 8 months after sensitization eyes will respond with an inflammatory reaction following the intravenous injection of fractions of the total antigen. 4. Repeated daily intravenous injections of a single antigen usually produce no reaction in the sensitized eye after the first few days. Injection of different antigens intravenously on succeeding days produces a continued sterile inflammatory process in the sensitized eye. After the total number of single antigens has been injected, repetition of these injections now fails to produce a similar response. Instead, the eye reaction is at a much lower level and the inflammatory response is manifested only to a few of the antigens injected intravenously. 5. Unless massive doses of antigen are used to desensitize, permanent desensitization of the eye has not occurred in animals which have been followed for at least 8 months. Animals may develop maximal eye responses following repeated intravenous injections of the same antigen, if sufficient time has elapsed between injections. Nevertheless, the eye reactions which can be elicited 6 or 7 months after sensitization are less intense than the initial responses. 6. The ability of the eye inflammation to light up following the intravenous injection of homologous antigen is not due to an initial tissue injury as proven by the fact that the reaction is specific and anterior chambers injured with typhoid vaccine, iodine, saline, glycerine, or albolene will not respond subsequently when the various proteins used for sensitization of the other eyes are injected intravenously. 7. It has been impossible to demonstrate sensitivity in the eye by the intravenous shocking route until at least the 5th day following introduction of the antigen into the anterior chamber. 8. The eye reaction can be produced by the subcutaneous as well as the intravenous injection of antigen. 9. Rabbits vary considerably in the intensity of the eye reaction which can be elicited in them, but only rarely was an animal found which failed completely to give a reaction.