bluuebunny/crossref_metadata_2025 huggingface dataset:
CELL TO CELL INTERACTION IN THE IMMUNE RESPONSE
Authors: Miller J. F. A. P. , Mitchell G. F.
Published: June 2004

An injection of viable thymus or thoracic duct lymphocytes was absolutely essential to enable a normal or near-normal 19S liemolysin-forming cell response in the spleens of neonatally thymectomized mice challenged with sheep erythrocytes. Syngeneic thymus lymphocytes were as effective as thoracic duct lymphocytes in this system and allogeneic or semiallogeneic cells could also reconstitute their hosts. No significant elevation of the response was achieved by giving either bone marrow cells, irradiated thymus or thoracic duct cells, thymus extracts or yeast. Spleen cells from reconstituted mice were exposed to anti-H2 sera directed against either the donor of the thymus or thoracic duct cells, or against the neonatally thymectomized host. Only isoantisera directed against the host could significantly reduce the number of hemolysin-forming cells present in the spleen cell suspensions. It is concluded that these antibody-forming cells are derived, not from the inoculated thymus or thoracic duct lymphocytes, but from the host. Thoracic duct cells from donors specifically immunologically tolerant of sheep erythrocytes had a markedly reduced restorative capacity in neonatally thymectomized recipients challenged with sheep erythrocytes. These results have suggested that there are cell types, in thymus or thoracic duct lymph, with capacities to react specifically with antigen and to induce the differentiation, to antibody-forming cells, of hemolysin-forming cell precursors derived from a separate cell line present in the neonatally thymectomized hosts.
CELL TO CELL INTERACTION IN THE IMMUNE RESPONSE
Authors: Nossal G. J. V. , Cunningham A. , Mitchell G. F. , Miller J. F. A. P.
Published: June 2004

Two new methods are described for making chromosomal spreads of single antibody-forming cells. The first depends on the controlled rupture of cells in small microdroplets through the use of a mild detergent and application of a mechanical stress on the cell. The second is a microadaptation of the conventional Ford technique. Both methods have a success rate of over 50%, though the quality of chromosomal spreads obtained is generally not as good as with conventional methods.

These techniques have been applied to an analysis of cell to cell interaction in adoptive immune responses, using the full syngeneic transfer system provided by the use of CBA and CBA/T6T6 donor-recipient combinations. When neonatally thymectomized mice were restored to adequate immune responsiveness to sheep erythrocytes by injections of either thymus cells or thoracic duct lymphocytes, it was shown that all the actual dividing antibody-forming cells were not of donor but of host origin. When lethally irradiated mice were injected with chromosomally marked but syngeneic mixtures of thymus and bone marrow cells, a rather feeble adoptive immune response ensued; all the antibody-forming cells identified were of bone marrow origin. When mixtures of bone marrow cells and thoracic duct lymphocytes were used, immune restoration was much more effective, and over three-quarters of the antibody-forming mitotic figures carried the bone marrow donor chromosomal marker.

The results were deemed to be consistent with the conclusions derived in the previous paper of this series, namely that thymus contains some, but a small number only of antigen-reactive cells (ARC), bone marrow contains antibody-forming cell precursors (AFCP) but no ARC, and thoracic duct lymph contains both ARC and AFCP with a probable predominance of the former. A vigorous immune response to sheep erythrocytes probably requires a collaboration between the two cell lineages, involving proliferation first of the ARC and then of the AFCP. The results stressed that the use of large numbers of pure thoracic duct lymphocytes in adoptive transfer work could lead to good adoptive immune responses, but that such results should not be construed as evidence against cell collaboration hypotheses.

Some possible further uses of single cell chromosome techniques were briefly discussed.
INTERACTIONS OF THE COMPLEMENT SYSTEM WITH ENDOTOXIC LIPOPOLYSACCHARIDE: CONSUMPTION OF EACH OF THE SIX TERMINAL COMPLEMENT COMPONENTS
Authors: Gewurz H. , Shin H. S. , Mergenhagen S. E.
Published: June 2004

Large amounts of each C'3, C'5, C'6, C'7, C'8, and C'9 were consumed when guinea pig serum was incubated with endotoxic lipopolysaccharide, zymosan, or preformed immune complexes. Since these C' components subserve several of the biological activities which follow the injection of endotoxins into experimental animals, these experiments support the hypothesis that certain biological effects induced by endotoxins may be mediated via the C' system, and may account for some of the known similarity in the reactivities evoked by endotoxins and immune complexes in vivo.
CELLS INVOLVED IN THE IMMUNE RESPONSE
Authors: Singhal Sharwan K. , Richter Maxwell
Published: June 2004

Cell suspensions of immune rabbit lymph nodes and spleen were capable of undergoing blastogenesis and mitosis and of incorporating tritiated thymidine when maintained in culture with the specific antigen in vitro. They did not respond to other, non-cross-reacting antigens. The blastogenic response obtained with immune lymph node cells could be correlated with the antibody synthesizing capacity of fragment cultures prepared from the same lymph nodes. Cell suspensions of immune bone marrow responded to non-cross-reacting antigens only whereas cell suspensions of immune thymus, sacculus rotundus, and appendix did not respond when exposed to any of the antigens tested.

On the other hand, neither fragments nor cell suspensions prepared from lymph nodes, spleen, and thymus of normal, unimmunized rabbits responded with antibody formation and blastogenesis when exposed to any of the antigens. However, normal bone marrow cells responded with marked blastogenesis and tritiated thymidine uptake. The specificity of this in vitro bone marrow response was demonstrated by the fact that the injection of a protein antigen in vivo resulted in the loss of reactivity by the marrow cell to that particular antigen but not to the other, non-cross-reacting antigens. Furthermore, bone marrow cells of tolerant rabbits failed to respond to the specific antigen in vitro. It was also demonstrated that normal bone marrow cells incubated with antigen are capable of forming antibody which could be detected by the fluorescent antibody technique. This response of the bone marrow cells has been localized to the lymphocyte-rich fraction of the bone marrow.

It is concluded that the bone marrow lymphocyte, by virtue of its capacity to react with blastogenesis and mitosis and with antibody formation upon initial exposure to the antigen, a capacity not possessed by lymphocytes of the other lymphoid organs, has a preeminent role in the sequence of cellular events culminating in antibody formation.
THE ULTRASTRUCTURE OF HUMAN THYROGLOBULIN
Authors: Bloth B. , Bergquist R.
Published: June 2004

Thyroglobulin molecules purified in a single step procedure by gel filtration were studied in the electron microscope using the negative staining technique. The molecule had the shape of a flexible helix with two turns. Its length was about 220 A and the maximal diameter of the coiled part of the molecule was estimated to be 110 A. The pitch varied between 40 and 50 A. Thyroglobulin molecules dried in sodium tungstosilicate on a carbon film as for electron microscopy retained their hemagglutination-inhibiting activity and 19S sedimentation constant when redissolved in physiological buffer.
CHEMOTAXIS OF MONONUCLEAR CELLS
Authors: Ward Peter A.
Published: June 2004

Chemotaxis of rabbit mononuclear cells was studied by the micropore-filter technique. Mononuclear cells obtained from mineral oil-induced peritoneal exudates respond chemotactically to rabbit serum treated with immune complexes or with streptokinase and plasminogen, to soluble factors produced by bacteria, and to lysates obtained from rabbit neutrophils.

The first chemotactic factor requires heat-labile factors in serum for its generation, but, once formed, the chemotactic factor is relatively heat stable. This factor has been compared with the complement-associated factor in serum, C'(5, 6, 7)a, that is chemotactic for neutrophils. The ability to generate the mononuclear cell chemotactic factor in serum that has been treated with potassium thiocyanate suggests that complement is not required. The position of the chemotactic factor in preparative electrophoresis and density-gradient ultracentrifugation indicates that on the basis of physical-chemical criteria, this factor is not C'(5, 6, 7)a.

The mononuclear cell chemotactic factor present in lysates of neutrophils sediments slowly in the ultracentrifuge and may be related, at least in part, to cationic peptides of lysosomal granules.

A study in the time course of the chemotactic response of mononuclear cells reveals that the response begins to level off after 4 or 5 hr. This is in sharp contrast to the time course for the chemotactic response of neutrophils, in which the reaction is complete within 1.5 hr.

Rabbit mononuclear cells obtained from a starch-induced peritoneal exudate respond to serum treated with plasminogen and streptokinase and to the soluble factor produced by bacteria, but no chemotactic response is elicited to serum treated with immune complexes. This indicates a functional difference between two populations of mononuclear cells. Rabbit alveolar macrophages respond poorly to all agents tested, although a weak chemotactic response to bacterial factors was found.

The requirement of a serine esterase in the mononuclear cell for the cell to respond chemotactically was defined by the use of organophosphorus inhibitors. Pretreatment of mononuclear cells with several series of phosphonates renders them unresponsive in the chemotactic system. The effect is similar for mononuclear cells responding chemotactically to activated serum and to the bacterial chemotactic factor. Several points of contrast with inhibition profiles obtained in the chemotactic system of the neutrophil suggest that, while the mononuclear cell requires a serine esterase for chemotactic responsiveness, this enzyme is different from the one previously defined in the neutrophil.
CROSS-REACTIONS AMONG GROUP A STREPTOCOCCI
Authors: Wiley Grove G. , Bruno Pauline N.
Published: June 2004

Strains of four streptococcal types, 33, 41, 43, 52, and a nontypable strain, Ross, cross-reacted in precipitin and bactericidal tests. The homologous reactions, which determined the type, afforded the major protection and developed promptly and regularly in the serum of rabbits during immunization. The associated cross-reactions, on the other hand, appeared in the serum of certain rabbits only, were often not as strong as the associated homologous reactions, and required for their presence a longer period of immunization than the homologous reactions.

Agar gel analysis of the homologous precipitin reactions revealed, as would be expected, reactions of serological identity, while those cross-reactions which were strong enough to test in this way formed bands of precipitate which joined with spur formation on the side of the homologous reaction.

These experiments and others referred to in the text suggest that cross-protection, as demonstrated in bactericidal tests, is sufficiently widespread to be a factor in streptococcal immunity, if a corresponding protection occurs in vivo. Thus, streptococcal infection with one of the cross-reacting strains might confer, in addition to strong homologous protection, a certain amount of cross-protection.
RECEPTORS FOR COMPLEMENT ON LEUKOCYTES
Authors: Lay Waltraut H. , Nussenzweig Victor
Published: June 2004

Sheep red blood cells sensitized by 7S, but not by 19S rabbit anti-Forssman antibodies, adhere and form rosettes on mouse macrophages and on a few monocytes and polymorphonuclear cells (PMN). When, however, C' factors from mouse serum are added to the antigen-19S antibody complex (EAC'), rosettes are formed on most mouse peritoneal macrophages and PMN and on a few monocytes. In addition EAC' also adheres to 10–25% of lymph node lymphocytes but not to thymus lymphocytes. EAC' prepared with 7S anti-Forssman antibodies has identical properties. The adherence of red cells induces an increase in the membrane activity of the leukocytes and causes injury to the red cells which rapidly become deformed and fragmented.

Adherence of EAC' occurs at 37°C and is minimal at 4°C. Probably only the first four C' components are involved in this phenomenon as mouse serum deficient in C'5 or rabbit serum, deficient in C'6 can be used as a source of C' components. Treatment of EAC' with EDTA does not modify its leukocyte-adherence properties. The adherence of EAC' to the leukocytes is not inhibited in the presence of serum.

The receptors for C' on macrophages, PMN, and monocytes differ from those found on lymphocytes. Rosette formation by EAC' on macrophages, PMN, and monocytes depends on divalent cations (Mg++) and can be reversed by Na3H EDTA, while adherence to lymphocytes is independent of these ions and occurs in the presence of 0.01 M Na3H EDTA. Both types of receptors for C' components are destroyed by trypsin treatment of the leukocytes, in contrast with the receptors for 7S antibodies on the same cells which persist after enzyme treatment.
IMMUNOLOGICAL STUDIES IN ULCERATIVE COLITIS
Authors: Lagercrantz R. , Hammarström S. , Perlmann P. , Gustafsson B. E.
Published: June 2004

The incidence and height of antibody titers to colon, assayed by indirect hemagglutination with a heat stable colon extract from germ free rats, is significantly higher in sera from patients with ulcerative colitis than in those from healthy controls or from patients with amebic liver abscess or dysentery. While sera from ulcerative colitis patients and controls are indistinguishable in regard to incidence and height of antibody titers to Forsman antigen, Staphylococcus aureus S 209, Clostridium difficile, and several common strains of E. coli, they have elevated titers and increased incidence of antibodies to a heat stable antigen of E. coli O14. Patients with amebic dysentery have normal titers of such antibodies.

Absorption of patients' sera with E. coli O14 antigen inhibits the colon directed hemagglutination reaction in approximately 30% of the cases tested. Likewise, the anti-E. coli O14 reaction can sometimes be inhibited with the colon extract. Other E. coli strains and other bacteria are inactive or have only weak inhibitory activity. Hemagglutination inhibition experiments show that germ free rat colon and E. coli O14 contain common structures, depicted by antibodies in the patients' sera. This pattern of reactivity closely resembles that seen in rats made autoimmune to colon by injection of newborn rabbit colon.

E. coli O14 is known to carry a heterogenetic antigen present in lower concentration (or activity) in most Enterobacteriaceae. Hemagglutination inhibition experiments with rabbit antisera to E. coli O14 suggest that the antigen common for E. coli O14 and colon is related to this heterogenetic antigen. The findings imply that this antigen, which is constantly present in low concentrations in the human colon, may give rise to anticolon antibody formation in ulcerative colitis through breakage of tolerance. Since this antigen is present in healthy individuals as well, additional factors are required to explain the induction of anti-colon autoimmunity in ulcerative colitis.
ENDOTOXIN AS ADJUVATOR TO THE TRANSPLANTATION OF A MOUSE MAMMARY TUMOR
Authors: Henderson James Stuart
Published: June 2004

The mouse mammary tumor MT 296 was used in a further series of experiments on the implantation of tumor, plated out in vivo, from suspensions of individual cells.

Lipopolysaccharide from S. typhosa was shown to exert an adjuvator influence. But this adjuvator, an endotoxin, had no direct effect on the suspended tumor cells, unlike the liver preparations previously reported.

Lipopolysaccharide from S. typhosa was shown to act on the host. It made the host's connective tissue expanses more susceptible to successful implantation by the tumor cells. It did this only if present at the time these connective tissue expanses were split. The increased susceptibility, caused by splitting the connective tissue expanses in the presence of lipopolysaccharide, declined quickly after 24 hr.

The structural changes wrought upon the connective tissues by splitting them in the presence of lipopolysaccharide are described. They show kinship to a Schwartzman reaction of the local type. Their possible role in the adjuvator effect on the plating of single cell suspensions of this tumor is discussed.
BONE MARROW AS SOURCE OF CELLS IN REACTIONS OF CELLULAR HYPERSENSITIVITY
Authors: Lubaroff David M. , Waksman Byron H.
Published: June 2004

The precise origin of cells infiltrating tuberculin skin reactions was studied with the technique of immunofluorescence. Thymectomized, irradiated Lewis rats were restored with bone marrow from allogeneic or F1 donors. They were passively sensitized to tuberculin by a subsequent transfer of Lewis lymph node cells and were given intradermal skin tests with tuberculoprotein. In 24 hr reactions the majority of cells were shown to be derived from the infused marrow. These results were the same regardless whether the lymphocyte transfer was performed on the day of irradiation and marrow injection or 7 days later. The cells in the tuberculin reactions, marrow, spleen, and lymph nodes not derived from the bone marrow were found to originate in the transferred lymph node cells. The relative percentages of marrow-derived and lymph node-derived cells in the tuberculin reactions remained the same during the 9–24 hr period following skin test.
IMMUNOLOGICAL INDUCTION OF INCREASED VASCULAR PERMEABILITY
Authors: Henson P. M. , Cochrane C. G.
Published: June 2004

Two mechanisms have been described whereby rabbit platelets in vitro may be induced to release their contained histamine by the reaction of antigen and antibody. Both processes require the participation of the complement system.

In the first, the adherence of platelets to particulate antigens such as zymosan or erythrocytes which have fixed complement through the third component was followed by histamine release. Plasma lacking C6 activity was fully active in this system.

In the second mechanism, the reaction of soluble antigen with antibody in the presence of plasma also caused release of histamine from platelets and platelet clumping was observed. The release process, which appeared to follow the adherence of platelets to the immune complex, required the action of C6 and perhaps the later-acting components. No evidence could be obtained that a soluble factor was produced which caused the release. Instead, an inhibitor of the release process was detected after incubation of antigen, antibody, and plasma.

Antibody preparations capable of giving complement-dependent PCA reactions in rabbits were also shown to induce the release of histamine from platelets in vitro.
THE SERUM BACTERICIDAL SYSTEM: ULTRASTRUCTURAL CHANGES IN NEISSERIA MENINGITIDIS EXPOSED TO NORMAL RAT SERUM
Authors: Swanson John , Goldschneider Irving
Published: June 2004

Exposure of meningococci to the bactericidal system of normal rat serum initiates a series of ultrastructural changes that accompany death of the organism. These morphological alterations consist of complement-dependent holes in the cell wall outer membrane, edema of the periplasmic space and cytoplasm, and accumulation of fibrillar material in and on the cell wall. Dissolution of the dense line and rupture of the cytoplasmic membrane also occur in meningococci exposed to normal rate serum. These latter two changes, however, are not seen in meningococci that are killed by bentonite-absorbed (lysozyme-deficient) serum, nor are they invariably present when other serum-susceptible organisms (N. catarrhalis and Herellea sp.) are treated with normal rat serum.

The pattern of development of edema in serum-treated meningococci suggests that the cell wall outer membrane, mucopeptide layer, and cytoplasmic membrane act synergistically to maintain osmotic equilibrium of the bacterium. Death of the bacterium seems related to alteration of permeability following injury to the outer membrane.

Holes are demonstrable, by negative straining, in the outer membrane of the meningococcal cell wall after exposure to the bactericidal effects of rat serum. The lesions are 110 A average greatest diameter and depend on the presence of antibody and complement for their formation. In addition, 82 A diameter holes are present in the walls of normal, untreated meningococci. The relation of complement-dependent holes to the smaller, naturally occurring holes is not known.
A NEW METHOD FOR THE ENUMERATION OF ANTIGEN-REACTIVE CELLS RESPONSIVE TO A PURIFIED PROTEIN ANTIGEN
Authors: Armstrong W. D. , Diener E.
Published: June 2004

A new technique for the enumeration of antigen-reactive cells (ARC) responsive to the polymer antigen of S. adelaide flagellin (POL) is described for two strains of mice. Foci have been shown to be antibody dependent, may be mimicked by IgM as well as IgG antibodies, and contain specific antibody-forming cells (AFC). The use of POL offers a system unencumbered by relatively high numbers of background foci which, when present, appear to be basically different from those found using the SRBC antigen.

The response of 1 antigen-reactive cell (ARC) focus/1 x 106 CBAT6T6 mouse spleen cells is linearly related to the injected number between 1 x 106–3 x 106 donor spleen cells and since 5% of injected cells remain in the spleen, there are an estimated 2400 ARC/spleen.

The number of ARC foci does not increase significantly after the 5th postantigen day, and by the 8th day the AFC progeny of ARC have reached the maximum mean of 280 AFC/ARC focus. In response to increasing antigen concentrations, an initial rise in the number of AFC as well as ARC is observed, resulting in a relatively constant AFC/ARC ratio. This suggests that the number of ARC stimulated determines the total number of AFC produced under these conditions rather than a variable mitotic rate of the ARC offspring. The main significance of this technique is that it will allow a study of the kinetics of the ARC in the primary and secondary immune response as well as in immunological tolerance.
TISSUE INTERACTION IN THE DEVELOPMENT OF THYMUS LYMPHOCYTES
Authors: Owen J. J. T. , Ritter M. A.
Published: June 2004

Thymic rudiments from chick and mouse embryos have been cultured in diffusion chambers on the chick chorioallantois. In this situation their supply of blood-borne stem cells is cut off. Although rudiments from older embryos become fully lymphoid under these circumstances, primordia from early embryos fail to develop any lymphoid cells. Early chick rudiments will however develop completely if they are grafted directly to the chorioallantois where they receive a vascular supply.

It is concluded that stem cells first enter the chick thymic anlage at between 6 and 7 days' incubation and the mouse thymic rudiment between 10 and 11 days' gestation. During the following few days of development there is a rapid inflow of stem cells to the rudiments. Since it is likely that stem cells, once they have entered the thymic primordium, are capable of only limited proliferation, it must be concluded that an inflow of stem cells continues once full lymphopoiesis has begun, although perhaps at a reduced rate.

Finally, the importance of the interaction between stem cell and organ rudiment to normal thymic development is discussed in relation to the pathogenesis of thymic anomalies.
THE FREQUENCY OF ANTIGEN-SENSITIVE CELLS IN TISSUE TRANSPLANTATION
Authors: Nisbet Norman W. , Simonsen Morten , Zaleski Marek
Published: June 2004

Graft-vs.-host (GVH) reactions were performed in chicken embryos by intravenous injection of adult chicken blood dilutions, and the result was scored by weighing the spleens as well as by karyological identification of host and donor metaphases. From the frequency of detectable signs of GVH reaction when low doses of donor cells were injected into hosts containing one foreign allele of the B locus it is concluded that 1–2% must be a minimum estimate for the frequency of antigen-sensitive cells of a given specificity in this system. It is not found possible to reconcile the findings with the strictest form of clonal selection which postulates a single receptor specificity per clone of antigen-sensitive cells.
PATHOGENESIS OF CHRONIC DISEASE ASSOCIATED WITH PERSISTENT LYMPHOCYTIC CHORIOMENINGITIS VIRAL INFECTION
Authors: Oldstone Michael B. A. , Dixon Frank J.
Published: June 2004

Mice infected shortly after birth with lymphocytic choriomeningitis (LCM) virus are not immunologically tolerant, although they carry the virus throughout life. These LCM carrier mice make anti-LCM antibody, which apparently complexes with viral antigen in the circulation and these complexes accumulate in the glomeruli.

LCM carrier mice of different strains vary significantly as to concentration of detectable infectious virus in their tissue, amount and time of appearance of anti-LCM antibody, and development of an associated chronic disease. The chronic disease consists primarily of glomerulonephritis, focal hepatic necrosis, and disseminated lymphoid infiltrations. LCM carriers of the SWR/J strain contain high tissue concentrations of virus, considerable anti-LCM antibody detectable in the glomeruli by 3 wk to 2 months of age and develop chronic disease within the first 2–3 months of life. In contrast, C3H strain LCM carriers contain 1/1000 as much infectious virus, less detectable anti-LCM antibody, and have not, over a 24 month observation period, developed any detectable disease. B10D2 old and new carrier mice with intermediate amounts of virus develop chronic disease during the latter half of the first year of life.

The pathogenesis of the glomerulonephritis of chronic LCM disease is apparently related to the formation of circulating virus-antibody complexes which are trapped in the glomerular filter. There is no evidence for direct glomerular injury by the virus nor for any autoimmune response by the host.
EFFECT OF ADRENALECTOMY ON BLOOD PRESSURE IN SALT-FED, HYPERTENSION-PRONE RATS
Authors: Iwai Junichi , Knudsen Knud D. , Dahl Lewis K. , Tassinari Lorraine
Published: June 2004

In adrenalectomized, genetically hypertension-prone rats, a high degree of correlation was found between evidence of functioning adrenal tissue and the development of salt hypertension. There is considerable evidence that some rats have the capacity to regenerate functioning adrenal cortical tissue from accessory glands and microscopic rests, sometimes in remote locations. Therefore, the criteria for continued absence of adrenal function after surgical adrenalectomy are critical. In this study we used three tests to validate the presence, or absence, of adrenal function: (a) a biochemical test, the quantitative, serial measurement of plasma glucocorticoids in individual rats; (b) a physiological test, the ability to survive a virtually sodium-free diet; and (c) the anatomical search for histological evidence of adrenal cortical tissue. Among those animals that developed hypertension after adrenalectomy, the correlation between plasma steroid concentration and blood pressure was statistically significant. We suspect that this correlation exists only during the period when cortical tissue is regenerating; it does not exist among intact animals with and without hypertension induced by salt.

It was concluded that some adrenocortical function is necessary for salt hypertension to develop. The evidence was insufficient to settle the question whether the action of corticosteroids is causative, or whether they play a supporting, although necessary, role for an extraadrenal hypertensinogenic factor to become manifest.
FLUID PHASE DESTRUCTION OF C2hu BY C1hu
Authors: Gigli Irma , Austen K. Frank
Published: June 2004

The fluid phase inactivation of C2hu by C1hu is markedly enhanced by the presence of C4hu. The enhancement is afforded by C1 inactivated C4hu, namely C4ihu, and requires the simultaneous presence of enzymatically active C1. Heterologous C4 of guinea pig origin protects C2hu from the inactivation by C1hu. Thus, in both the fluid phase and on the cellular intermediate, C4hu is essential to the specific action of C1hu on C2hu. It is possible that C4i alters C2 so as to present a more suitable substrate to the C1 enzyme or that C4i acts on the C1 to uncover a specificity for native C2.
CYSTIC FIBROSIS OF THE PANCREAS
Authors: Danes B. Shannon , Bearn Alexander G.
Published: June 2004

103 individuals from 16 families with cystic fibrosis and 87 individuals without family history of cystic fibrosis have been studied using the methods of cell culture. Skin fibroblast cultures derived from 19 affected children, and fibroblast cultures from 11 different organs obtained at autopsy from two affected children, showed cellular metachromasia. The morphological appearance and the intracellular mucopolysaccharide content enabled these cultures to be divided into two distinct classes. Class I had discrete cytoplasmic metachromatic vesicles and a mucopolysaccharide content similar to that observed in normal fibroblasts. In class II the metachromasia was present in both vesicles and granules and was evenly distributed throughout the cytoplasm. The mucopolysaccharide content of these cells was markedly increased.

The cultures derived from the parents, presumed heterozygotes, and other members of each family showed cells with the same type of metachromasia as that demonstrated by the propositus. These data strongly suggest that cystic fibrosis is not a homogeneous entity and, moreover, can be caused by homozygosity of genes at two distinct loci.

The recognition of cytoplasmic abnormalities in skin fibroblasts derived from affected individuals and heterozygous carriers for cystic fibrosis should facilitate genetic and biochemical studies on the heterogeneity of this inborn error of metabolism.
EFFECTS OF NEOMYCIN AND PENICILLIN ADMINISTRATION ON MUCOSAL PROLIFERATION OF THE MOUSE SMALL INTESTINE
Authors: Khoury Kenneth A. , Floch Martin H. , Herskovic Teodoro
Published: June 2004

The effects of an oral neomycin and penicillin regimen on intestinal bacteriology and on morphology and function of the small intestine of mice were investigated. Quantitative and qualitative stool cultures on selective media of the treated animals revealed only growth of yeast organisms. The treated animals developed enlargement of the ceca with fluid contents and watery stools, resembling characteristics of germfree animals.

Radioautography with tritiated thymidine revealed an increased epithelial cell migration rate in the mice treated with the antibiotics for 3 to 5 wk. A slight increase in villus height was also noted. The treated male mice showed greater variance than the treated females in epithelial cell migration rates.

Histochemical staining reactions showed a decrease in nonspecific esterase and in NADH dehydrogenase activity in the proximal gut of the antibiotic animals. Stains of distal gut and those for acid and alkaline phosphatase, NADPH dehydrogenase, lactic dehydrogenase, and succinic dehydrogenase were similar to the controls. A slight increase in sucrase activity and a slight decrease in lactase activity in the antibiotic animals was observed in contrast to control animals. Germfree mice, however, had greater sucrase and lactase activity. Transport of L-methionine was slightly reduced in the distal segment of the treated animals.

Since the direction of these changes is away from the intestinal state observed in germfree animals, they are probably the result of the direct action of the antibiotics on the gut.
THE RENAL HANDLING OF HEMOGLOBIN
Authors: Bunn H. Franklin , Esham William T. , Bull Robert W.
Published: June 2004

The glomerular filtration of hemoglobin (α2ß2) was studied under conditions in which its dissociation into αß dimers was experimentally altered. Rats receiving hemoglobin treated with the sulfhydryl reagent bis(N-maleimidomethyl) ether (BME) showed a much lower renal excretion and prolonged plasma survival as compared with animals injected with untreated hemoglobin. Plasma disappearance was also prolonged in dogs receiving BME hemoglobin. Gel filtration data indicated that under physiological conditions, BME hemoglobin had impaired subunit dissociation. In addition, BME hemoglobin showed a very high oxygen affinity and a decreased rate of auto-oxidation.

Glomerular filtration was enhanced under conditions which favor the dissociation of hemoglobin into dimers. Cat hemoglobin, which forms subunits much more extensively than canine hemoglobin, was excreted more readily by the rat kidney. The renal uptake of 59Fe hemoglobin injected intra-arterially into rabbits varied inversely with the concentration of the injected dose.
THE RENAL HANDLING OF HEMOGLOBIN
Authors: Bunn H. Franklin , Jandl James H.
Published: June 2004

The fate of small doses of isotopically labeled isologous hemoglobin was studied in the rat. When haptoglobin depleted animals were given 2.0 mg of 59Fe hemoglobin intravenously, nearly half was trapped by the kidneys. Kidney 59Fe activity disappeared slowly over several weeks. Whatever iron was lost from the kidneys was largely reutilized. In contrast, the porphyrin of hemoglobin absorbed by the kidneys appeared to be rapidly catabolized, since 5 hr after the injection of 14C or 59Fe heme-labeled hemoglobin only a small fraction was recovered as hematin. Likewise, after injection of globin-labeled hemoglobin, rapid disappearance of kidney protein activity indicated that the absorbed globin was readily catabolized in situ.
STUDIES OF PPLO INFECTION
Authors: Simberkoff Michael S. , Thorbecke G. Jeanette , Thomas Lewis
Published: June 2004

Extracts of five arginine-utilizing mycoplasmas inhibit PHA-induced lymphocyte mitosis, while extracts of five glucose-utilizing mycoplasmas do not. Evidence is presented supporting the view that the inhibitory factor is the enzyme arginine deiminase. This enzyme inhibits the reactions of human lymphocytes to antigens as well as PHA, and the secondary production of antibody by rabbit lymph node fragments in vitro. Addition of enzyme to the cells several days after the initial mitotic or antigenic stimulus reduces, but does not abolish, further cellular activity. The production of serum proteins by hepatoma cells is totally unaffected by the mycoplasmal extract. It is concluded that arginine is an essential amino acid for the small lymphocyte, but not for the transformed cell nor for a number of other cell types. Suggestive evidence has been obtained that other enzymes similarly affect lymphocyte reactions.
THE EFFECT OF CYTOTOXIC AGENTS ON THE PRIMARY IMMUNE RESPONSE TO LISTERIA MONOCYTOGENES
Authors: Tripathy S. P. , Mackaness G. B.
Published: June 2004

Four drugs, representing four different categories of cytotoxic agents, were studied for their effect on the immune response to Listeria infection in mice. The development of the host's immune response is revealed by a progressive change in the slope of the bacterial growth curve in spleen and liver. It has its onset at 24 hr in untreated mice, but in the presence of effective immunosuppression the organism multiples uninterruptedly until the animal dies from overwhelming infection. When administered as single injections at the time of infection, cyclophosphamide, vinblastine, and azathioprine all produced an effective immunosuppression, characterized by continuous bacterial multiplication.

Methotrexate was also immunosuppressive, but unlike the others its effects were reversible. They could be sustained, however, by further treatment.

Studies of the time-response relationship indicated that cyclophosphamide was highly active over a broad time-span ranging from 2 days before infection to 4 days after infection. Vinblastine on the other hand was maximally active when given on the day of infection, while methotrexate had its greatest effect when given 48 hr after infection. These differences indicate that these three drugs act on different cell populations involved in the host's immune response. The effects observed have been discussed in relation to what is known of the modes of action of the drugs tested.

An observation of interest was the phenomenon of enhanced immunity in animals treated with cyclophosphamide or vinblastine 7–11 days before, and with methotrexate 4 days before infection; reactive hyperplasia of lymphoid tissue following withdrawal of drug was again advanced as an explanation for the occurrence of this paradoxical effect.

The experimental model employed is simple, requiring only routine bacteriological facilities and minimal equipment. It seems to offer a useful means of assessing the immunosuppressive activity of drugs and of determining the time-course of their action; it could also be of value in the screening of anticancer agents.
CELLS INVOLVED IN THE IMMUNE RESPONSE
Authors: Abdou Nabih I. , Richter Maxwell
Published: June 2004

The antigen-reactive cells in normal rabbit bone marrow could be isolated from a suspension of marrow cells by passage of the cells through an antigen-sensitized glass bead column. The cells which passed through the column were deficient in antigen-reactive cells directed to the antigen used to sensitize the glass beads, whereas the cells eluted from the column could transfer antibody-forming capacity to irradiated recipients only with respect to the specific sensitizing antigen. Separation of the bone marrow antigen-reactive cells could not be achieved by passage of the cells through nonsensitized glass bead columns or in the presence of excess free antigen in the column. Cells which were retained by, and later eluted from, the antigen-sensitized glass bead columns were mostly small mononuclear cells, whereas cells which passed through the columns were morphologically similar to the original unfractionated bone marrow cell suspension. The data indicate the presence of an antibody or antibody-like structure, with defined immunological specificity, on the surface of the normal bone marrow antigen-reactive cell.
THE MECHANISM OF ACTION OF ANTI-LYMPHOCYTE SERUM
Authors: Lance Eugene M.
Published: June 2004

Studies designed to gain insight into the mechanism of action of the active component of antilymphocyte serum were carried out using an antibody eluate prepared from the IgG fraction of anti-lymphocyte serum by absorption and subsequent elution from thymocyte membrane. The resulting antibody eluate was labeled with radionuclide tracer to determine the fate of the antibody in vivo. The result indicated that anti-lymphocytic antibodies are eliminated from recipients extremely rapidly. The mechanism for this rapid clearance appears to depend upon the absorption of antibody molecules onto lymphocyte surfaces and the subsequent clearing and degradation of the antibody-lymphocyte complexes by the reticuloendothelial system.

Distribution studies confirm that the major site of antibody-lymphocyte interaction occurs in the periphery with relatively little penetration of antibody within lymphoid organs. Radioautographic studies showed that the pattern of localization within lymphoid and other organs is confined to rather specific areas. These observations are believed to offer strong support for the notion that anti-lymphocyte serum achieves its immunosuppressive effect by bringing about a selective ablation of the population of recirculating lymphocytes.
THE ANAMNESTIC ANTIBODY RESPONSE TO TYPE III SPECIFIC PNEUMOCOCCAL POLYSACCHARIDE
Authors: Paul William E. , Benacerraf Baruj , Siskind Gregory W. , Goidl Edmond A. , Reisfeld Ralph A.
Published: June 2004

Rabbits immunized with killed type III pneumococci respond to anamnestic challenge by type specific polysaccharide (S III) with the synthesis of anti-S III antibody if a long interval is allowed to elapse between primary and secondary immunization. A study of the anti-S III antibody produced early and late in the immune response revealed no change in molecular class, banding pattern of dissociated light chains, or S III binding characteristics as measured under equilibrium conditions or by study of dissociation kinetics utilizing radioiodinated p-OH-benzyl-S III.

Sequential booster injections of S III into rabbits primarily immunized with whole organisms 8 or 9 months earlier led to a progressive decrease in the number of animals showing successful anamnestic responses and in the magnitude of those responses.

It is concluded that S III depletes the antigen sensitive cell population in the secondary response largely because of its limited ability to stimulate sustained proliferation by such cells.
TRANSFER OF EXPERIMENTAL AUTOIMMUNE THYROIDITIS BY SERUM FROM THYROIDECTOMIZED DONORS
Authors: Nakamura R. M. , Weigle W. O.
Published: June 2004

When rabbits were injected with 10.0 mg rabbit thyroglobulin in complete Freund's adjuvant, the earliest thyroid lesions were seen on day 5 and uniformly severe thyroid lesions were seen by day 14; these observations were not significantly different from the thyroid lesions observed at 1 and 2 months post-immunization. Pooled sera were obtained from immunized, thyroidectomized, and nonthyroidectomized donors on various days and transferred to normal recipient rabbits in different experiments. Successful transfer of thyroid lesions was seen when serum containing early antithyroglobulin antibody obtained from thyroidectomized donor animals at various times after immunization was injected into normal recipients in a sequential manner. Immunofluorescent studies of recipient thyroid glands showed focal fixation of rabbit γ-globulin and ß1C complement in thyroid follicles. When purified antibody to rabbit thyroglobulin obtained from thyroidectomized donor sera was transferred sequentially as above, significant thyroid lesions were seen in recipient rabbits. In contrast, no thyroid lesions were seen in recipient animals injected with rabbit sera containing late antithyroglobulin antibody from thyroidectomized donors or hyperimmune sera from guinea pigs.

No thyroid lesions were seen in recipient animals injected either with sera from donors given complete adjuvant without thyroglobulin or with globulin fraction of pooled sera containing early antithyroglobulin antibody obtained on various days from nonthyroidectomized donors. Similarly, rabbits rendered unresponsive to guinea pig γG-globulin and periodically injected with guinea pig anti-rabbit thyroglobulin showed no thyroid lesions.
THE MITOTIC POTENTIAL OF FIXED PHAGOCYTES IN THE LIVER AS REVEALED DURING THE DEVELOPMENT OF CELLULAR IMMUNITY
Authors: North Robert J.
Published: June 2004

Fixed macrophages in the liver of the mouse undergo mitosis in large numbers during an infection with Listeria monocytogenes. The macrophage mitotic response always precedes the expression of efficient host immunity to infection. It is suggested that macrophage proliferation is an important event in the development of host immunity to L. monocytogenes.
MATURATION OF HEMOLYSIN-PRODUCING CELL CLONES
Authors: Saunders George C.
Published: June 2004

Investigations of the induction period of an in vitro hemolysin response to sheep erythrocyte antigen revealed the following:

1. After antigen stimulation precursors of plaque-forming cells rapidly maturate to the point of hemolysin production.

2. Initial maturation probably occurs in the absence of cell division.

3. After initial maturation, a latent period of about 12 hr occurs before the first doubling of PFC.

4. At least the first three cell doublings are synchronous, with a generation time of 7–8 hr.

5. Synchronous cell division implies that all precursor cells are at the same point in the cell cyde when they are initially stimulated.
IDIOTYPY OF RABBIT ANTIBODIES
Authors: Oudin Jacques , Michel Mauricette
Published: June 2004

The idiotypic patterns detected in the anti-Salmonella typhi serum of one given bleeding have been sought in other bleedings of the same rabbit. Idiotypic patterns carried by antibodies of the second bleeding are not always found in those of the first bleeding. Bifurcated precipitation zones in gels (double diffusion in cells) have been observed in the reaction of several anti-idiotypic sera with the sera of the first and second bleedings of three rabbits, and apparently indicate that idiotypic patterns which were carried by the same molecule in the first bleeding were carried by two separate molecules in the second bleeding. In a comparison of idiotypy of two anti-S. typhi serum samples of one rabbit, the collection of which had been separated by a very long interruption of the immunization, all the idiotypes detected in the early bleeding were also detected in the late bleeding; several idiotypes were detected in the serum of the late bleeding and not in that of the early bleeding.

Idiotypy has been observed and compared in IgM and in IgG antibodies. There are, in each of these two classes, antibodies that are not distinguished from antibodies of the other class by anti-idiotypic sera, and consequently that carry the same idiotypic patterns.

A comparison has been made between the idiotypy of the two kinds of antibodies which, in anti-S. typhi sera, are precipitated and not precipitated by the polysaccharide. Certain idiotypic patterns are carried only by the antibodies of the first kind. There are also idiotypic patterns which are carried by antibodies of these two kinds, among which certain anti-idiotypic antibodies do not discriminate.

Possible cellular implications of observations made on idiotypy at different stages of the immunization of one individual have been discussed.
INDUCTION AND RECALL IN CONTACT SENSITIVITY
Authors: de Sousa Maria A. B. , Parrott Delphine M. V.
Published: June 2004

The cellular events in the ear skin and draining lymph node during the induction of contact sensitivity to 2-ethoxy methylene-5-oxazolone (oxazolone) have been studied in three strains of mice.

The principal findings in the skin during the first 24 hr were invasion of polymorphs and destruction of pilosebaceous units, in both intact and thymectomized mice. Subsequently, the dermal cellular infiltrate increased and there was acanthosis of the epidermis. No lymphocytes were seen in the dermis or penetrating the epidermal basal cell layer in thymectomized mice.

During the first 24 hr in the draining node, polymorphs and macrophages bearing a pigment with staining properties similar to melanin were seen in the marginal and medullary sinuses, in intact and thymectomized mice. Major differences, however, were revealed during 2–4 days when massive proliferation of large pyroninophilic blast cells occurred in the thymus-dependent area of the nodes from intact mice only.

On testing, there was a prompt, measurable increase in ear thickness only in intact mice. This increase reached a peak at 24 hr — typical of a delayed type reaction. At testing, the ears from intact mice showed epidermal vesiculation and a considerable dermal cellular infiltrate with a substantial number of lymphocytes. This was in contrast with the completely quiescent appearance of the ear skin of thymectomized mice.

Finally, we have discussed the use of the mouse as an experimental tool for studying contact sensitivity and have analyzed the role of the thymus-derived lymphocyte and the site where it becomes sensitized, in the light of current theory on the origin of cells and site where sensitization takes place in cell-mediated reactions.