bluuebunny/crossref_metadata_2025 huggingface dataset:
The molecular requirements for chemotactic attraction of leucocytes by proteins. Studies of proteins with synthetic side groups
Authors: Wilkinson P. C. , McKay I. C.
Published: February 2007

Various small organic molecules were covalently linked to human serum albumin (HSA) and other proteins, and the products were tested for their ability to attract human peripheral blood neutrophil leucocytes in vitro. It was found that many substances, when attached to HSA, could increase its chemotactic activity. The chemotactic activity of the conjugates was not specifically inhibited by the presence of the conjugated substance in free solution. Thus it seems unlikely that chemotactic recognition of structurally altered proteins depends on a close stereospecific bonding between a cell surface receptor and the side group on the protein.

In general, non‐polar side groups such as tosyl, propionyl and butyryl groups made the protein more chemotactic, than did polar side groups such as picryl, p‐sulphobenzylazo, guanidyl and carbamoylmethyl groups. We suggest that the chemotactic recognition of structurally altered proteins by neutrophils depends on hydrophobic bonding between non‐polar groups on the protein and those on the cell surface. Ionic groups would also be necessary for activity if the protein had many surface non‐polar groups: if there were no coulombic repulsions the non‐polar groups would become concealed by adhering to one another.
The occurrence of antigenic determinants common to flagella of different salmonella strains
Authors: Langman R. E.
Published: February 2007

Five Salmonella proteins, hitherto regarded as antigenically distinct, have been shown to share common antigenic determinants. It is proposed that the flagellar (H) antigens be designated as either Hv (variable) or Hc (common); in these terms the Kauffmann‐White schema describes the Hv determinants only. Antibodies to Hc determinants could be demonstrated by precipitation reactions detected by the Ouchterlony gel diffusion technique and by passive hemagglutination assays.

To account for the finding that antibody to Hc determinants is not detected by bacterial immobilization assays it was proposed that Hv determinants are distributed along the flagella so that antibody can bind with two sites on the one flagella, thus disrupting the wave motion; in contrast, antibody to Hc determinants can only bind to one flagella at a time and therefore acts as a crosslink and does not cause immobilization.
Carrier specificity and the allogeneic effect in mice
Authors: Rajewsky K. , Roelants G. E. , Askonas Brigitte A.
Published: February 2007

The findings of several investigators suggest thatin vitrosoluble T cell factors, non‐antigen specific, can replace helper T cells in the induction of antibodies. The present experiments set limits to the possible range of action of such factors in thein vivoinduction process.

Murine cells primed with a hapten‐carrier complex could not be efficiently stimulated to produce anti‐hapten antibody by injection of the carrier alone or together with the hapten on an unrelated carrier. This reconfirms the requirement of physical union between hapten and carrier for efficient secondary stimulation.

Even under conditions of presumably maximal production of non‐antigen specific T cell product, namely in a graft‐versus‐host reaction, there was no evidence for the systemic appearance of such a factor. A graft‐versus‐host reaction could substitute for specific helper cells only when the hapten‐primed cells were the targets in the cellular reaction and did not belong to the aggressor cell population.

The allogeneic effect was shown to be T cell‐dependent and to correlate with reactivity in mixed lymphocyte reaction. In most instances, efficient allogeneic stimulation of the secondary anti‐hapten response required the presence of both allogeneic aggressor cells and hapten on a heterologous carrier.

Very low numbers of allogeneic cells (around 104T cells per host) sufficed to abrogate carrier specificity almost completely. This suggests that some type of amplification mechanism is involved in mediating the allogeneic effect.

The results support the concept thatin vivo, efficient antibody induction relies on cell‐cell interaction mediated by antigen bridging or by allogeneic recognition.
Thymus cells are inducible to osteogenesis
Authors: Friedenstein A. J. , Lalykina Klavdia S.
Published: February 2007

Guinea pig and rabbit thymus cells showed a marked capacity for bone formation under the influence of transitional epithelium and decalcified bone matrix and were much superior in that respect to all the other populations of lymphoid and hemopoietic cells studied. Diffusion chambers containing the reacting cells together with inductors of osteogenesis were used.
Genetic control of immune response in mice to derivatives of multichain polyproline differing in the optical configuration of component amino acids
Authors: Mozes Edna , Sela M. , Mcdevitt H. O.
Published: February 2007

The immune response of two inbred mouse strains, DBA/1 and SJL, to eight synthetic polypeptides of the general formulae, poly(Tyr, Glu)‐poly Pro—polyLys and poly(Phe, Glu)‐polyPro—polyLys, differing in the optical configuration of the amino acids composing the macromolecules, was compared. SJL mice responded predominantly to the polyPro—polyLys moieties of either optical configuration in the eight enantiomorphs. In both strains of mice tested, immunization with polypeptides of the formula poly(Tyr, Glu)‐polyPro—polyLys leads to antibodies specific for the polyproline moiety mainly. DBA/1 mice are low responders to 701, poly(LTyr, LGlu)‐polyLPro—polyL Lys, whereas they elicit a good immune response to polyDPro—polyDLys when immunized with 713, poly(DTyr, DGlu)‐polyDPro—polyDLys, and with 711, poly(LTyr, LGlu)‐polyDPro—polyDLys. It is of interest that mice of the DBA/1 strain responded well to 715, poly(DTyr, DGlu)‐polyLPro—polyLLys with antibodies specific to poly‐L‐proline. Since this polymer is metabolized slowly and incompletely similarly to antigens composed exclusively of D‐amino acids, it is likely that the slow metabolism affects the ability of this mouse strain to respond to this immunogen.

DBA/1 mice responded predominantly to the (LPhe, LGlu) region when immunized with 702, poly(LPhe, LGlu)‐polyLPro—polyLLys. Since they are high responders to (LPhe, LGlu) and to polyDPro—polyDLys, they elicited antibodies to both immunopotent regions of 712, poly(LPhe, LGlu)‐polyDPro—polyDLys. Although DBA/1 mice are high responders to (LPhe, LGlu) they did not respond to D‐amino acid analogues of these peptides, and immunization of this mouse strain with 714, poly(DPhe, DGlu)‐polyDPro—polyDLys, and with 716, poly(DPhe, DGlu)‐polyLPro—polyLLys, elicited anti‐polyproline antibodies only.

The results of this study show that the optical configuration of the amino acids composing an antigen plays an important role in the genetic control of the ability to respond to the immunogens investigated.
Synthesis of surface immunoglobulin by lymphocyte leukemia cells in vitro
Authors: Moroz C. , Shalmon Lea , Hahn J.
Published: February 2007

The synthesis of surface immunoglobulin by chronic lymphocytic leukemia cells was studied in short‐term culture using radioactive amino acids and sugars. The membranal location of the nascent immunoglobulin was evidenced by the difference in the electrophoretic pattern of the membranal and cytoplasmic proteins, and by the absence of nascent surface immunoglobulin in cultures containing chain‐specific antibody. The nascent surface immunoglobulins comprised 7 S IgM and a large excess of free light chains of lambda type. Anti‐ü‐chain serum blocked the nascence of membranal IgM, while λ‐type as well as K‐type anti‐L‐chain sera blocked the nascence of both membranal IgM and free light chains. Radioactive glucosamine and mannose were not incorporated into the nascent surface immunoglobulin. There was no secretion of nascent proteins into the culture medium.
Immunoglobulins on the surface of sheep lymphocytes I. Class and cellular distribution
Authors: Ey P. L.
Published: February 2007

Sheep immunoglobulins were purified and rabbit antisera specific for μ and γ heavy chains and light chains prepared. Lymphocytes obtained by cannulation of the efferent duct of the sheep popliteal node were labeled in vitro with 125I‐anti‐Ig antibodies. The percentage of lymphocytes labeling for μ, γ or light chains, was, in each case, approximately 20 – 25 %, but cells were most heavily‐labeled by anti‐μ chain and least by anti‐γ chain antibodies. As the sum of the proportions of cells labeling for μ and γ chains (total, 40.6 %) greatly exceeded that labeling for light chains (20.4 %), it was concluded that labeled cells had both γ and μ chains exposed on their surfaces, probably as IgG and IgM.

Lymphocytes were titrated against anti‐μ or anti‐γ chain sera and means (± 1 standard deviation) of 19 200 ± 8200 molecules IgG and 14 700 ± 9300 molecules IgM (19 S, or 73 600 ± 46 800 molecules 8 S) were calculated to be exposed on the surface of each cell labeled by anti‐Ig antibodies.

It is suggested that those cells from the sheep which bound anti‐Ig antibodies were bone marrow‐derived lymphocytes as has been found in the mouse.
Cell‐mediated immune response in vitro. IV. Metabolic studies on cellular immunogenicity
Authors: Wagner H.
Published: February 2007

The relationship between the immunogenicity of living allogeneic cells and their metabolism was investigated in an in vitro cytotoxic allograft system. Cortisone‐resistant CBA mouse thymocytes (H‐2k) were activated in vitro against mitomycin C‐treated Balb/c spleen cells (H‐2d), and the resultant cytotoxic response was assayed against 51Cr labeled H‐2d target cells. Inhibitors which function at the level of transcription (actinomycin D) and oxydative phosphorylation (antimycin A) were applied to block selectively the allogeneic (stimulating) cells. While 5 × 106 – 7 × 106 mitomycin C‐treated Balb/c spleen cells induced optimal cytotoxic immune responses in vitro, actinomycin D or antimycin A‐treated allogeneic cells were 25‐fold less immunogenic. This quantitative defect in immunogenicity could be repaired by increasing the number of immunogenic cells. Cells killed by freeze‐thawing or heating had almost lost their immunogenicity. Thus, there appeared to be a relationship between active cell metabolism and the immunogenicity of allogeneic cells.

When the concentration of immunogenic cells was raised to 20 × 106, it was found that mitomycin C‐treated cells stimulated the system suboptimally, suggesting the possibility of tolerance induction. However, actinomycin D‐ or antimycin A‐treated cells actually induced a good immune response at this cell concentration. The theoretical implications of the results are discussed.
Specific characterization of isoelectrofocused immunoglobulins in polyacrylamide gel by reaction with 125I‐labeled protein antigens or antibodies
Authors: Keck K. , Grossberg A. L. , Pressman D.
Published: February 2007

A method is described for the visualization and characterization of isoelectrofocused antibodies, immunoglobulins in general, and many other proteins in thin plates of polyacrylamide gel. The novel aspect of the method is that the protein bands after electrofocusing are immobilized in the gel by treatment with glutaraldehyde after soaking with 18 % sodium sulfate solution. Such treatment is shown not to affect the binding activity of antibodies. Antibody bands are located by treatment of the plates with 125I‐labeled specific antigen. Other proteins can be located by 125I‐labeled specific antibody. The bands thus located are visualized by radioautography. The immobilization of the focused proteins allows the excess radiolabeled reagent macromolecules to be washed out of the plates without disrupting the focused pattern. Under the conditions described, background radioactivity and nonspecific pickup by other serum proteins in the plates are negligible.

Examples of the use of the method are given for anti‐4‐azonaphthalene‐1‐sulfonate and anti‐4‐hydroxy‐5‐iodo‐3‐nitro‐phenacetyl antibodies, using the 125I‐labeled hapten‐specific antigens, and for a mouse IgG myeloma protein, MOPC 195, using 125I‐labeled goat anti‐mouse IgG antibody.

Extension of the method would allow differences between proteins to be characterized by appropriate 125I‐labeled antibodies. Thus immunoglobulins could be characterized by 125I‐labeled antibodies specific for immunoglobulin markers (e.g. Ig class, L chain type, etc.).

The advantages of this method over other methods include the ease and generality of using radioiodine labeled macromolecules – especially antibodies – as compared to the restriction imposed by other methods which require small labeled ligands. The method can be extended to the use of fluorescent‐labeled proteins and radiolabeled polysaccharide antigens as locators.
Immunoglobulin levels of sera of genetically thymusless (nude) mice
Authors: Bloemmen J. , Eyssen H.
Published: February 2007

The immunoglobulin levels of the various mouse immunoglobulin classes in sera of genetically thymusless (nu/nu) mice and their littermate controls with thymus (nu/+) were determined. The amounts of IgM in nu/nu and control mice were equal, but the amounts of IgA, IgG1, IgG2a and IgG2b in nu/nu mice of 10 weeks were, respectively, 14, 3, 27 and 14 % of those in control mice. Our results indicate that the presence of an intact thymus is not needed for the production of IgM, but seems to play an important part in the production of IgA and IgG.
Somatic mutation and the origin of antibody diversity. Clonal variability of the immunoglobulin produced by MOPC 21 cells in culture
Authors: Cotton R. G. H. , Secher D. S. , Milstein C.
Published: February 2007

A method has been developed for the screening of immunoglobulins produced by large numbers of individual clones of the mouse myeloma (MOPC 21) in tissue culture. Clones grown in agar were incubated in the presence of radioactive amino acids and applied directly onto polyacrylamide plates for isoelectric focusing. A discrete number of well‐defined immunoglobulin bands were then detected by autoradiography. The following variations between different clones were observed:

1. Variation in relative intensity of the different bands.

2. Total absence of immunoglobulin bands.

3. Variation in the position of the bands along the pH gradient.

When a mutant clone exhibiting this third type of variation was purified by subcloning, only the mutant pattern was observed. Thus, the cells appear to express only a single allele, although they are known to be aneuploid.

We consider the third type of variation to be relevant in the evaluation of the contribution of somatic mutation to the diversity of antibody structure.
The extent of diversity of anti‐hapten antibodies in inbred mice: anti‐NIP (4‐hydroxy‐5‐iodo‐3‐nitro‐phenacetyl) antibodies in CBA/H mice
Authors: Kreth H. W. , Williamson A. R.
Published: February 2007

The number of different anti‐NIP (4‐hydroxy‐5‐iodo‐3‐nitro‐phenacetyl) molecules which inbred CBA/H mice can make was estimated. Spleen cell transfer has been used and the isoelectric spectra of antibodies present in the sera of recipient mice determined. Comparison between the sets of monoclonal antibody spectra deriving from different donor mice gave a repeat frequency for individual spectra. This frequency permitted a statistical estimate (see Addendum) of the minimum number of different anti‐NIP molecules potentially capable of being elicited in CBA/H mice. The most likely value of this pool is 8000 with 98 % probability that pool exceeds 3000 and 95 % probability that pool is less than 30 000.
Immunization against abolition of transplantation tolerance*
Authors: Ramseier H.
Published: February 2007

Attempts were made to break the tolerance of murine transplantation antigens of strain A in CBA mice by adoptive transfer of: 1) normal, syngeneic (CBA) lymphoid cells, or 2) moderate doses of sensitized, syngeneic (CBA anti‐A) lymphoid cells followed later by high doses of CBA anti‐A cells. Both kinds of attempt failed. Tolerance could be broken when a large enough dose of sensitized cells was given. Prior “immunization” of CBA mice tolerant to A with small doses of CBA spleen cells protected against doses of sensitized cells otherwise successful in breaking tolerance.

Determinations of anti‐recognition structure (anti‐RS) antibody and of alloantibody activities in sera of tolerant mice, of mice which were successful in defending tolerance, and of mice in which tolerance could be abolished revealed an important role for anti‐RS antibodies. No clear role could be assigned to alloantibodies present in the same sera. The function of anti‐RS antibodies was shown by the failure of spleen cells from immunized tolerant mice to recognize tolerated antigen. This suggests that anti‐receptor antibodies are active in neutralizing receptors of responding lymphoid cells emerging from a stem cell reservoir.
Binding and release of lymphocytes by hapten‐derivatized nylon fibers
Authors: Kiefer H.
Published: February 2007

We have studied the binding of lymphocytes to hapten‐derivatized nylon fibers at different temperatures and found that cells bound at 4 °C for more than 12 h can be released quantitatively from the fiber at temperatures above 22 °C within 2 h in a fully viable state. Experiments with lymphocytes bound to 2,4‐dinitrophenyl (DNP)‐derivatized nylon mesh and then stained with Rhoda‐mine‐labeled rabbit anti‐mouse Ig antibodies showed that the cells had capped and that the cap was directed toward the fiber in all cases. This result, as well as the temperature dependence of binding and release, suggest that capping of the lymphocytes is involved in their release from the fibers.
Affinity and Ig classes of anti‐hapten antibodies in carrier‐preimmunized rats
Authors: Hurme M. , Kontiainen Sirkka , Seppälä I. J. T. , Mäkelä O.
Published: February 2007

Priming rats with the carrier protein 7 – 43 days before injecting them with a conjugate of hapten NIP [(4‐hydroxy‐5‐iodo‐3‐nitrophenyl)acetyl] and the protein accelerated the anti‐hapten response: peak titers were reached sooner, the replacement of IgM by IgG antibody happened earlier and the affinity of anti‐NIP was higher in carrier‐primed than in non‐carrier‐primed rats. The acceleration effect on peak titers was short‐lasting. By day 9 the anti‐hapten titers in non‐carrier‐primed animals had reached those of the carrier‐primed animals. The difference in affinities gradually decreased, but was still demonstrable 28 days after immunization with the conjugate.
Antigenicity of dinitrophenyl polyproline
Authors: Gurari Dalia , Ungar‐Waron Hana , Sela M.
Published: February 2007

Several poly‐L‐proline batches, as well as their dinitrophenyl derivatives, were synthesized and characterized. Their immunogenicity in rabbits, guinea pigs and mice was investigated, and the specificity of the antibodies formed was determined. For sensitive detection of anti‐polyprolyl antibodies, polyprolyl bacteriophage T4 was prepared. Polyproline was in this case attached to the bacteriophage via 1,3‐difluoro‐4,6‐dinitrobenzene.

Dinitrophenyl poly‐L‐proline is immunogenic in rabbit and guinea pig, but essentially nonimmunogenic in the mouse. No genetic differences in the antibody response were found either in strains 2 and 13 of guinea pigs, where both strains were good antibody producers, or in several strains of mice, all of which were essentially unresponsive. Poly‐L‐proline, considered nonimmunogenic in rabbits by previous criteria, led to a low but significant antibody response, when tested with polyprolyl bacteriophage.
Role of a rigid polyproline spacer inserted between hapten and carrier in the induction of anti‐hapten antibodies and delayed hypersensitivity
Authors: Ungar‐Waron Hana , Gurari Dalia , Hurwitz Esther , Sela M.
Published: February 2007

The synthesis and characterization are described of a hapten‐protein conjugate in which carrier [BSA or the synthetic multichain polymer, poly‐L‐(Tyr, Glu)‐poly‐DL‐Ala–poly‐LLys, (T,G)‐A–L] and hapten 2,4‐dinitrophenyl (DNP) are separated by the rigid peptide chain of an L‐proline oligomer. This compound has been used to study the carrier effect in an attempt to distinguish between the cooperation and local environment theories which have been put forward to explain it.

Rabbits were given a primary injection of the antigen DNP‐poly‐Lprolyl‐BSA (bovine serum albumin). The secondary response was followed in vitro after stimulation with antigens composed of hapten coupled to carriers in the presence or absence of a polyproline spacer, and compared to secondary stimulation with the homologous antigen. Antibody production was monitored by thymidine incorporation, by inactivation of modified bacteriophage and by hemolytic plaque formation. Experiments in delayed hypersensitivity were also carried out using the same antigens. In both systems it was found that the insertion of the inflexible spacer molecule between hapten and carrier did not abolish the carrier effect. These results lend support to the cooperation hypothesis as explanation of the carrier effect.
Specificity of IgA cold agglutinins: anti‐Pr1
Authors: Roelcke D.
Published: February 2007

Monoclonal cold agglutinins have usually been found to be IgM(k) and to be directed against the I/i antigens.

Three IgA(k) cold agglutinins were tested for antibody specificity. Based on the following criteria, they were shown to be directed against the Pr1antigen of the Pr1/Pr2/Praantigen system. The antigens reacting with the IgA(k) cold agglutinins were inactivated by protease and treatment of human red cells with neuraminidase and were demonstrable on human erythrocyte glycoproteins. The terminal sugar of the determinant group of these antigens is N‐acetylneuraminic acid (NANA), which is not involved in I/i, but also in Pr2antigenicity. The antigen activity of these antigens and the Pr2and I antigen activities were distributed to different glycopeptide fractions obtained from red cell glycoproteins by ficin cleavage and Sephadex G‐50 separation. Periodate oxidation of red cell glycoproteins, which causes a shortening of the polyhydroxy side chain at C‐6 of C‐9 NANA to a C‐7 and/or C‐8 NANA derivative, resulted in inactivation of these antigens, while the I antigen remained unchanged and the Pr2antigen was increased 16–64‐fold. Dog red cells gave no or diminished reactions with the three IgA(k) cold agglutinins, in contrast to increased reaction with anti‐Pr2. Based on the different reactions of the IgA(k) cold agglutinins with dog red cells, a Pr1heterogeneity Pr1h/Pr1dwas demonstrated.

In spite of the determination of the Pr1h/Pr1d/Pr2and the MN antigens by NANA, both antigen systems were shown to be unrelated. Blocking of free ε‐lysine amino groups by acetylation or blocking of the NANA carboxyl groups by amidation of red cell glycoproteins resulted in MN inactivation, while the antigens reacting with the IgA(k) cold agglutinins (like Pr2) remained unaffected.

The anti‐Pr1specificity of IgA cold agglutinins and the predominance of anti‐I/‐i specificity of IgM cold agglutinins was discussed with respect to inter‐relations between immunoglobulin classes and antibody specificities of cold agglutinins.
Tumor immunity to murine plasma cell tumors. II. Essential role of T lymphocytes in immune response
Authors: Rouse B. T. , Röllinghoff M. , Warner N. L.
Published: February 2007

Spleen cells, thoracic duct lymphocytes and adherent peritoneal exudate cells from mice immunized to syngeneic plasma cell tumors were capable of transferring specific protective immunity to these tumors. Pre‐treatment of these cells with anti‐Θ serum or anti‐lymphocyte serum, but not with anti‐ΘK serum, effectively abolished the ability of lymphoid cells to transfer this immune response. These studies demonstrate that T cells are essential in the adoptive transfer of immunity. Furthermore, immunization of athymic (nude) mice to plasma cell tumors was not achieved. Attempts to activate normal macrophages in vitro with sensitized lymphoid cells were unsuccessful. Admixture of immune lymphoid cells and target tumor cells to unrelated tumor cells did not inhibit the growth of the unrelated tumor cells. It is proposed that the immune response to syngeneic tumor associated antigens of plasma cell tumors not only involves essential sensitization of T lymphocytes, but that activated cytotoxic T lymphocytes are directly responsible for inhibiting target tumor growth.
Role of T lymphocytes in the pathogenesis of experimental autoimmune encephalomyelitis
Authors: Lennon Vanda Alice , Byrd W. J.
Published: February 2007

A dichotomy between clinical and histological signs of experimental autoimmune encephalomyelitis (EAE) was demonstrated in neonatally thymectomized (NNTx) rats. Although neonatal thymectomy prevented subsequent induction of clinical EAE in Lewis rats challenged as young adults with syngeneic basic protein (BP) of myelin, there was a 50 % incidence of histological EAE. Both cellular and humoral immune responses to BP were reduced in the NNTx rats. However, the presence of a cell‐mediated response to BP (MIF release) was statistically associated with the occurrence of histological EAE.

Diminished preimmunization blood lymphocyte counts and impairment of the response of spleen cells to phytohemagglutinin were associated with reduced antibody responses. This suggests that collaboration between T and B cells may be necessary for production of antibody to BP. The relationship between antibody and clinical EAE is not clear and warrants further investigation.
Cellular events in protein tolerant inbred rats. I. The fate of thoracic duct lymphocytes and memory cells during tolerance induction to human serum albumin
Authors: Bell E. B. , Shand F. L.
Published: February 2007

(AS2 x AS)F1 hybrid rats were made completely or partially tolerant to an adjuvant challenge of human serum albumin (HSA) by prior treatment with fluid HSA (tolerogen). Single doses (10 μg to 200 mg) of fluid HSA failed to elicit a detectable immune response. However, rats made partially tolerant by repeated small doses (10 – 100 μg) synthesized significant quantities of antibody in response to the treatment.

Completely unresponsive rats (100 mg fluid HSA 3 x/week for 8 weeks) contained no detectable antigen‐binding antibody in their serum and no plaque‐forming cells (PFC) in their spleens. Thoracic duct lymphocytes from these animals failed to respond when adoptively transferred to irradiated, bone marrow restored recipients and did not inhibit the response of equal numbers of normal thoracic duct cells. Normal lymphocytes responded normally in completely tolerant rats, whether the recipients were irradiated or not. Cells from the bone marrow of tolerant rats were able to restore HSA responsiveness to irradiated rats within weeks of repopulation.

Partial tolerance was characterized by a reduction in PFC per spleen, a diminished antigen‐binding capacity (ABC) in the serum, a rapid decline in ABC with time and inhibition or abolition of a memory response. Analysis of the dosage requirements for HSA‐tolerance failed to support the concept that tolerance was induced at two separate dosage thresholds of antigen, i.e. “high” and “low” zone tolerance. To the contrary, the degree of unresponsiveness increased continuously in proportion to the dose of antigen and the number of injections, implying that tolerance induction can be explained by a single mechanism at all doses.
In vivo and in vitro immunological cross‐reactions between basic encephalitogen and synthetic basic polypeptides capable of suppressing experimental allergic encephalomyelitis
Authors: Webb Cynthia , Teitelbaum Dvora , Arnon Ruth , Sela M.
Published: February 2007

A significant extent of immunological cross‐reactivity has been demonstrated between the basic encephalitogenic protein of bovine origin and several synthetic amino acid copolymers which have suppressive effect on experimental allergic encephalomyelitis (EAE). This cross‐reactivity has been conclusively established on the cellular level, both in vivo by means of delayed hypersensitivity skin tests and in vitro using transformation of sensitized lymphocytes, as measured by incorporation of radioactive thymidine. The in vitro experiments have been conducted with lymph node cells from guinea pigs of both random bred and inbred strains, and on spleen and lymph node cells from rabbits. Definite cross‐reactivity was observed between the basic encephalitogen and all the synthetic copolymers which were previously shown effective in suppression of EAE, whereas ineffective copolymers or unrelated proteins did not show any cross‐reactivity. In the case of strain 2 guinea pigs and rabbit lymph node cells the cross‐reactivity in vitro was manifested by direct cross‐stimulation of the lymphocytes, whereas in random‐bred or strain 13 guinea pigs and rabbit spleen cells, the cross‐reaction was detected only by means of specific inhibition of the homologous stimulation by the heterologous antigen.

A limited extent of cross‐reactivity was observed on the humoral level as well; antibodies provoked in guinea pigs against the synthetic copolymer Cop 1 cross‐reacted in the passive cutaneous anaphylaxis assay with the bovine basic encephalitogen.
Rosette‐forming cell and cytophilic antibody
Authors: Liacopoulos P. , Couderc J. , Bleux C. , Birrien J. L. , Bach J. F.
Published: February 2007

Immunization of mice with two non‐cross‐reacting erythrocytes results in the appearance of double rosettes. Injection of sera from doubly immunized animals into normal mice also results in double rosette appearance when spleen cells are examined after rosette formation and incubation at 0 °C for 1 h. However, passive rosettes entirely disappear after overnight incubation at 4 °C or after incubation for 1 h at 37 °C, whereas active ones persist in significant numbers. These treatments therefore provide an accurate discrimination between cells passively sensitized by cytophilic antibody and cells actively forming antibody.

Removal of cells adhering to plastic surfaces from spleen cells of doubly immunized animals provokes a decrease of simple rosettes and a disappearance of doubles among non‐adherent cells.

When adherent cells are tested for existence of hemolytic plaque‐forming cells (PFC), it was found that PFC also adhere. Conversely, the non‐adherent cell population is severely depleted of such cells. Moreover, double PFC, again found in initial cell populations, were recovered with adherent cells but were absent in the final non‐adherent population.
Abrogation of immune tolerance to protein antigens in tolerant rabbits and mice by transfer of allogeneic spleen cells in small doses
Authors: Nachtigal D. , Zan‐Bar I.
Published: February 2007

When small doses of allogeneic spleen cells are injected into rabbits or mice made tolerant to either human or bovine serum albumin, about half the treated animals will escape from tolerance. The effective doses in rabbits are 2 x 106 to 5 x 106 cells per animal and in mice 104 to 106 cells per animal. Larger doses, around 25 x 106 cells in rabbits, are ineffective.

It is suggested that by transferring sufficiently small doses of allogeneic cells the threshold of the recipient's rejection mechanism can be circumvented.
Effét de la Rifamycine SV sur la réponse immunitaire induite in vitro
Authors: Thierry R. C.
Published: February 2007

Mouse peritoneal cells producing antibodies against sheep red cells were used to study the effect of Rifamycin SV on an in vitro induced immunological response by the localized hemolysis technique.

At a concentration of 100 μg/ml, Rifamycin SV has a greater inhibiting effect on lymphocytes than on macrophages. Lymphocytes are still clearly inhibited by 10 μg/ml, but the same concentration acting on macrophages causes an increase in the number of plaque‐forming cells. One thousand μg/ml Rifamycin SV completely suppresses antibody production, whereas 1 μg/ml has no detectable action.

The effect of Rifamycin SV on incorporation of radioactive amino‐acids and uridine was also studied. We found that it has a weak effect on protein synthesis but inhibits synthesis of RNA; this inhibition is stronger in the absence of antigen. Furthermore, protein synthesis by these cells seems to be controlled by long‐lived ribonucleic acids.
Quenching of tryptophan fluorescence in rabbit and bovine IgG
Authors: Hill R. J.
Published: February 2007

IgG, Fab fragments and poly‐D, L‐alanylated IgG prepared from normal rabbit sera show a decrease in fluorescence on titration down from pH 9 to 3. A qualitatively similar behavior is seen in a preparation of normal bovine IgG in both the native and heat‐denaturated state. This decrease in fluorescence is believed to be due to collisional quenching of tryptophan fluorescence by protonated amino acid residues. Since tryptophan is believed to be an integral part of the antigen‐binding site, these findings open the possibility of using tryptophan fluorescence to follow conformational changes in the antibody molecule.
Analysis of antibody populations to oligo‐alanine determinants. II. Kinetic characterization of allotypic subpopulations
Authors: Weiler Ivan Jeanne , Weiler E. , Dittrich B. , Sprenger Renate
Published: February 2007

Antibodies to the determinant oligo‐D‐alanine can be characterized by the speed of their association and dissociation with multivalent antigen. The antigen is present on the surface of sheep red blood cells (SRBC) as dense clusters, formed by poly‐alanylated rabbit antibodies against SRBC. Anti‐D‐ala IgG molecules attached to such “receptors” sensitize SRBC for subsequent lysis by complement, provided they are complexed with anti‐allotype antibodies (“facilitation”). Kinetic curves for the association reaction (at 5 °C) or for the dissociation reaction (at 35 °C) indicate whether antibody populations are heterogenous or homogenous in their kinetic properties.

Even when mice are of the same genetic constitution, antisera from individual mice may show up to 30‐fold differences in association velocities.

The use of anti‐allotype antibodies as facilitating reagents allows the separate kinetic analysis of allotypically defined antibody subpopulations within heterozygous animals. Such subpopulations were found to be distinct and independent with respect to association‐dissociation characteristics.
The capacity of macrophage components to inhibit anti‐macrophage serum activity
Authors: Schroit A. J. , Geiger B. , Gallily Ruth
Published: February 2007

Anti‐macrophage serum (AMS) was shown to lose part of its activity after incubation with macrophages. Both the cytotoxic and phagocytic inhibition properties of AMS decreased significantly after 12 h incubation with macrophages.

Free and/or AMS‐bound macrophage antigens were found to be shed from the cell surface into the culture medium during incubation of macrophages with AMS.

Labeled macrophage components were demonstrated to bind specifically to AMS, suggesting that the loss of AMS activity was mainly due to the blocking of antimacrophage antibody combining sites by macrophage antigens.
Cytophilic antibody: correlation of its distribution with activation of basophils and macrophages
Authors: Ferrarini M. , Munro A. , Wilson Anne B.
Published: February 2007

Immunoglobulin determinants on the surface of guinea pig basopils were detected using mono‐ and divalent antiglobulin reagents conjugated with fluorscein isothiocyanate. The fluorescent pattern obtained with monovalent antiglobulin was always that of uniformly stained rings. Divalent antiglobulin agglutinated immunoglobulin determinates to form small fluorescent sports subsequently collected at one end of the cell (“cap” formation). The formation of caps is temperature‐dependent and is inhibited by sodiu azide and cytochalasin B.

When basophils were incubated at 37 °C with divalent antiglobulin, morphological changes occurred, indicating that they had been activated. The same effect could be obtained with monovalent Fab́, but at least one hundred times more of it was required than the corresponding F(ab́)2.

Cytophilic antiboidies on the surface of guinea pig peritoneal macrophages were also detected by immunofluorescence. In this case again, divalent antiglobulin reagents agglutinated the determinants to form small fluorescent sports. Cap formation was not observed and pinocytosis started as the first aggregates were formed. It was also found that cytophilic antibodies were not stable on the macrophages' surface and were easily eluted at 37 °C. A reaction with cross‐linking agents (divalent antiglobulin or antigen) stabilized the cytophilic antibody and pinocytosis was initiated.
Generation of chemotactic activity by immune complexes carrying clustered or nonclustered C&42horbar; sites
Authors: Opferkuch W. , Snyderman R. , Borsos T.
Published: February 2007

Sensitized cells (EA) bearing different numbers of &42horbar; sites were tested for their ability to generate chemotactic activity from C‐EDTA. From the results it can be shown that:

the amount of chemotactic activity generated parallels the number of &42horbar; sites bound to the cell surface,

all &42horbar; sites clustered around a single hemolytic site are enzymatically active as far as generation of chemotactic activity is concerned, and,

no difference can be demonstrated with IgG or IgM antibodies
Immune responses in vitro. ii. mixed leukocyte interaction in a protein‐free medium
Authors: Katz‐Heber Ellen , Peck A. B. , Click R. E.
Published: February 2007

Addition of mercaptoethanol to the culture medium has permitted a mixed leukocyte interaction (MLI) to be achieved with mouse spleen cells in a chemically defined protein‐free medium. Results which confirm those obtained in culture medium supplemented with heterologous sera are: 1) the maximum incorporation of isotope occurred between 60 and 72 h of culture; 2) the index of stimulation was 3–15; 3) the number of responder and stimulator cells required for maximum incorporation was 3 x 106 – 4 x 106 and 4) immunization resulted in a higher level of incorporation that occurred earlier than the maximum response of normal cells. Other results obtained confirm previous reports which are in apparent conflict: i.e., 1) stimulation of parental strains (P1) by F1 hybrids (P1 x P2) may be either equal to or one‐half that induced by the other parental strains (P2); and 2) F1 hybrids may or may not be stimulated by the parental strains. These latter two results were found to be dependent on the strains used. Other advantages of the system are discussed in terms of the elimination of artifacts that occur in medium containing serum and the usefulness in examination of molecular events occurring early in immune reactions.
Precursor cells specific to sheep red cells in nude mice. estimation of frequency in the microculture system
Authors: Quintáns J. , Lefkovits I.
Published: February 2007

A dilution scheme for bone marrow‐derived (B) cells has been applied to estimate the frequency of precursor cells specific to sheep red cells (SRC) present in the spleens of nude mice.

B cells were titrated over a range of 0.2 x 105 – 1.8 x 105 spleen cells of nude mice per microculture in the presence of a constant number of allogeneic spleen cells. Data from 6 limiting dilution experiments are presented. It is argued that only the B cells are being diluted out. The frequency of precursor cells specific for SRC was calculated by means of Poisson statistics. The estimates of these frequencies are in the range 0.9 x 10‐5 – 2 x 10‐5.
in vitro complementation experiments with nude mice. i. the allogeneic effect in the antibody response to sheep red cells
Authors: Lefkovits I.
Published: February 2007

Complementation experiments were carried out in which allogeneic spleen cells were added to the spleen cells from nude mice. Evidence is presented that anti‐Θ sensitive T cells are responsible for the complementation. Experiments are described in which the anti‐sheep red cell response is followed as a function of the number of allogeneic spleen cells used to supplement the nude spleen cell cultures. With increasing numbers of allogeneic cells, optimum complementation is observed and then, a rapid drop of the response as the number of allogeneic cells is increased further. As few as 20 x 103 allogeneic spleen cells are able to complement 20 x 106 nude spleen cells to give a significant response. A delay in the addition of allogeneic cells up to 24 h does not change the kinetics of the response.
Hapten‐specific tolerance: unresponsiveness in the t celldepleted population
Authors: Hamilton J. A. , Miller J. F. A. P.
Published: February 2007

Complete and specific suppression of the anti‐NIP [(4‐hydroxy‐5‐iodo‐3‐nitro‐phenyl)acetyl] indirect plaque‐forming cell response to NIP‐fowl IgG was obtained in mice by pretreatment with NIP‐coated syngeneic erythrocytes. The complete suppression occurred within 5 to 7 days and was evident after adoptive transfer of spleen cells into irradiated mice. The state of unresponsiveness was reversed, not by thymus cells, but by supplementing with T cell‐depleted normal spleen cells. This suggests that the unresponsiveness occurred at the level of the B lymphocyte population.
In vitro induction of tumor specific immunity: requirements for T lymphocytes and tumor growth inhibition in vivo
Authors: Röllinghoff M. , Wagner H.
Published: February 2007

Cortisone‐resistant thymocytes, spleen cells, thoracic duct lymphocytes, peritoneal exudate cells and peripheral blood lymphocytes of BALB/c mice were immunized in vitro against syngeneic HPC‐108 plasma cell tumor cells. Cocultivation of spleen lymphocytes together with HPC‐108 cells generated the highest cytotoxic immune response in comparison to other lymphocyte sources. Cytotoxicity was tested in a 6 hour 51Cr release assay using HPC‐108 cells as target cells.

The use of AKR anti‐θ C3H serum indicated that thymus‐derived (T) lymphocytes are essential to the initiation phase of the immune response to plasma cell tumor cells. Furthermore, evidence is presented that the cytotoxic effector cells in the in vitro tumor immune response are T lymphocytes.

Spleen cells activated in vitro against HPC‐108 tumor cells were shown to specifically prevent tumor growth from simultaneously injected HPC‐108 cells in irradiated recipient mice.
Stimulation of DNA synthesis in mouse lymphoid cells by polyanions in vitro. I. Target cells and possible mode of action
Authors: Diamantstein T. , Vogt W. , Rühl H. , Bochert Gudrun
Published: February 2007

The effect of dextran sulfate on [3H]dThd incorporation in lymphoid cells was investigated. The polyanion activated DNA synthesis in spleen and bone marrow cells of normal mice. The highest rate of activation was detected in spleen cells of athymic (nude) mice; the ratio of [3H]dThd incorporation was higher in TxBM spleen cells of thymectomized, lethally irradiated and bone marrow protected mice than in spleen cells of normal donors. Thymus cells of normal mice could not be stimulated, but a slight response was obtained in cortisone‐resistant thymus cells. A synergetic effect was found in normal thymus cells using a combination of dextran sulfate and phytohemagglutinin (PHA). In contrast, lipopolysaccharide and PHA showed no synergetic effect.

The possible mode of action of polyanions as de‐repressors of DNA synthesis is discussed.
Surface immunoglobulin of myeloma cells
Authors: Bevan M. J.
Published: February 2007

Mouse myeloma cells have been stained in suspension at 0 °C or at room temperature with fluorescein‐labeled rabbit anti‐mouse immunoglobulin sera (F‐RAMIG) or with unlabeled RAMIG followed by fluorescein‐labeled goat anti‐rabbit immunoglobulin sera (F‐GARIG). Tissue culture adapted murine myeloma cells 5563 (IgG2aκ) have been used most extensively in these studies. 95–100 % of the cells show surface fluorescence at both temperatures after staining with F‐RAMIG or with RAMIG followed by F‐GARIG. At 0 °C the staining is diffuse over all the surface, at room temperature the staining is much brighter and patchy. The nature of the H and L‐chains at the surface of the myeloma cell is shown to parallel the type of secretory product. Thus, 5563 cells stain brilliantly with anti‐IgG and anti‐κ chain sera, but not at all with anti‐μ or anti‐α chain sera, which respectively stain TEPC 183 (IgMκ) and MOPC 47A (IgAκ) cells brilliantly. Also, MOPC 315 cells (IgAλ) do not stain with antisera against 5563 myeloma protein, but do stain with antisera against MOPC 47A and MOPC 104E myeloma proteins.

Immediately after staining at 0 °C or room temperature, the fluorescence is located over the whole surface. However, if the stained cells are incubated at 37 °C in tissue culture medium, the stain gradually localizes to one pole of the cell such that by 5 h, about 50 % of the cells show staining localized to one hemisphere. The presence of the RAMIG‐antigen complex seemed to have little or no effect on the rate of secretion of myeloma protein during a 5 h period. The results are discussed in relation of the findings of others and to the significance of the surface immunoglobulin of myeloma cells.
The failure to transfer the NZB disease following implantation of NZB thymus‐containing chambers
Authors: Tuffrey Maureen , Kingman Jean , Barnes R. D.
Published: February 2007

The role of the thymus in the disease of the NZB still remains to be clearly defined. Upon the basis of Coombs' positive reactions occurring in normal strains of mice following the implantation of thymus‐containing diffusion chambers, previous workers suggested the primary role of a subcellular thymus causal factor in the disease of the NZB. We have been unable to confirm these findings in neonatally and adult thymectomized or intact mice of two other strains in spite of being implanted with NZB thymus‐containing chambers.
Isoelectric spectra of different classes of anti‐erythrocyte antibodies
Authors: Phillips Jennifer M. , Dresser D. W.
Published: February 2007

A method is described for visualizing isoelectric spectra of antibody against foreign erythrocytes. Different classes of immunoglobulins may be examined separately. After electrofocusing, anti‐SRBC bands are detected by overlaying the gel with molten agarose containing SRBC and anti‐globulin sera for different antibody classes. On addition of complement, bands of specific lysis appear, which are recorded photographically.

Samples of sera from mice immunized with SRBC were electrofocused and bands of direct lysis and of γG2 — γG2a — developed lysis are shown. As the antigen dose is increased, more bands of γG2a anti‐SRBC antibody are detectable. The increase in the numbers of bands over the dose range used approximates to the increase seen in PFC over the same dose range.

This method is quick and easy to perform and can also be used to detect antibody spectra against proteins, haptens and other antigens which can be linked to indicator red blood cells.
Ability of the T cell‐replacing polyanion dextran sulfate to trigger the alternate pathway of complement activation
Authors: Hadding U. , Dierich M. , König W. , Limbert M. , Schorlemmer H. U. , Bitter‐Suermann D.
Published: February 2007

Dextran sulfate (DS) consumed C3 in C4 deficient guinea pig serum. This temperature‐dependent reaction required Mg++ ions and could therefore be blocked by EDTA. Isolated C3 was not influenced by DS, but serum factors were required for C3 consumption. The C3 proactivator as well as C3 were converted to their activated state by DS in guinea pig and human serum, as revealed by immunoelectrophoretical analysis. DS generated anaphylatoxin activity in serum. It is concluded that DS activates C3 via the alternate pathway of complement activation. This potency of the polyanion might serve as a tentative explanation for its T cell‐replacing effect in an antibody‐forming system, which was reported by Diamantstein and his coworkers.