bluuebunny/crossref_metadata_2025 huggingface dataset:
Lymphocytes bearing Fc receptors for IgE. V. Effect of tunicamycin on the formation of IgE-potentiating factor and IgE-suppressive factor by con A-activated lymphocytes.
Authors: Yodoi J , Hirashima M , Ishizaka K
Published: December 2022

Attempts were made to induce the formation of soluble factors having affinity for IgE (IgE-binding factors) by activated T cells. Normal rat mesenteric lymph node (MLN) cells were cultured in 1 microgram/ml Con A for 48 hr or in 10 microgram/ml Con A for 72 hr, and Con A-activated lymphocytes were incubated with rat IgE. It was found that IgE induced an increase in the proportion of Fc epsilon R(+) cells in the Con A-activated cells and formation of IgE-binding factors. However, the nature of IgE-binding factors formed by the cells were different depending on the concentration of Con A for activation. Thus, IgE-binding factors formed by 10 microgram/ml Con A-activated cells had a high affinity for lentil lectin and enhanced the IgE-forming cell response of DNP-OA primed cells. In contrast, the majority of IgE-binding factors formed by 1 microgram/ml Con A-activated cells failed to bind to lentil lectin and suppressed the IgE response. Induction of Fc epsilon R by IgE on Con A-activated cells was prevented by tunicamycin, which inhibits protein glycosylation. This antibiotic did not prevent the IgE-induced formation of IgE-binding factors but changed the nature of the factors formed by 10 microgram/ml Con A-activated cells. The majority of IgE-binding factors formed in the presence of tunicamycin lacked affinity for lentil lectin and Con A, and suppressed, rather than enhanced, the IgE response. The nature and amount of IgE-binding factors formed by 1 microgram/ml Con A-activated cells was not affected by tunicamycin. The results suggest that glycosylation of IgE-binding factors during their biosynthesis is an important step in determining their biologic function.
Activation and suppression of graft-vs-host reactions by cyclophosphamide.
Authors: Wander R H , Hilgard H R
Published: December 2022

We have investigated the effects of a single injection of cyclophosphamide (CY) on the graft-vs-host (GVH) reactivity of spleen cells from BALB/c mice. Six days after CY injection, spleen cells showed a broad range of reactivity when tested for their capacity to produce splenomegaly in young F1 hybrid recipients. Eight and 10 days after CY treatment, their capacity to produce GVH splenomegaly was markedly reduced. We also investigated the effects of untreated and CY-treated BALB/c spleen cells on the local GVH reaction that occurs when BALB/c lymph node pieces are grafted onto F1 host kidneys. Untreated BALB/c spleen cells augmented the local GVH reaction, whereas day 8 CY-treated BALB/c spleen cells reduced the local reaction. Day 6 CY-treated BALB/c spleen cells marginally reduced the local response and elicited considerable host splenomegaly. Day 6 CY-treated F1 hybrid spleen cells stimulated the local GVH reaction and did not produce splenomegaly. We interpret our results to indicate that CY treatment leads to a) the appearance on day 6 of a BALB/c cell population with increased GVH reactivity, b) the emergence of a suppressive mechanism by day 8 that results in reduced GVH reactivity, and c) an alteration at day 6 in F1 lymphoid cell antigenicity that enables these cells to stimulate an ongoing GVH reaction. We discuss our results in terms of the similar effects that CY has on GVH-reactive and autoreactive cell populations, and examine the possibility that the mechanisms that underlie GVH reactivity and autoreactivity are similar in that both involve imbalances of immunoregulatory cell populations..
Specificity of OKT4+ cytotoxic T lymphocyte clones.
Authors: Krensky A M , Clayberger C , Reiss C S , Strominger J L , Burakoff S J
Published: December 2022

OKT4+, HLA-DR-specific CTL were cloned by limiting dilution, and two clones were evaluated. One clone, B8, specifically recognized DR6 antigens, whereas another cloned, C6, recognized an Ia-like determinant on some DR 3, 5, and 6 target cells. Both clones were OKT3+, OKT4+, and OKT8-, and their cytolysis could be blocked by OKT3 and OKT4, but not OKT8, antibodies. A panel of monoclonal antibodies that recognize DR molecules blocked target cell recognition by these OKT+ CTL. Although cloned B8 recognized a DR6-specific determinant, clone C6 appeared to recognize a supratypic determinant that may be common to some DR molecules (e.g., MT2) or possibly another human Ia-like antigen (e.g., SB). The availability of OKT+ CTL clones should help to dissect the Ia-like antigens recognized by human T cells.
Structural and numerical variations of the carbohydrate moiety of immunoglobulin G.
Authors: Mizuochi T , Taniguchi T , Shimizu A , Kobata A
Published: December 2022

Oligosaccharide patterns obtained from human IgG by hydrazinolysis were fairly constant from sample to sample. In contrast, a variety of oligosaccharide patterns were obtained from IgG myeloma proteins. Structural analysis of each oligosaccharide by sequential exoglycosidase digestion and methylation studied indicated that the different patterns of IgG myeloma proteins are produced by different degrees of incompleteness of the formation of a single sugar chain: Sia alpha 2 leads to Gal beta 1 leads to 4GlcNAc beta 1 leads to 2 Man alpha 1 leads to 6(Sia alpha 2 leads to Gal beta 1 leads to 4GlcNAc beta 1 leads to Man alpha 1 leads to 3)(GlcNAc beta 1 leads to 4)Man beta 1 leads to 4GlcNAc beta 1 leads to 4(Fuc alpha 1 leads to 6)GlcNAcOT.
Biosynthesis of a structurally abnormal C2 complement protein by macrophages from C2-deficient guinea pigs.
Authors: Goldberger G , Cole F S , Einstein L P , Auerbach H S , Bitter-Suermann D , Colten H R
Published: December 2022

In order to characterize a genetic deficiency of C2 in guinea pigs, production of C2 by peritoneal macrophage cultures derived from four normal, four heterozygous deficient, and four homozygous deficient animals was measured functionally and immunochemically after metabolic labeling with 35S-methionine. Macrophage monolayers from homozygous deficient animals failed to secrete hemolytically detectable C2 up to 74 hr in culture. A single cell hemolytic plaque assay also failed to demonstrate any functional C2 production by cells from homozygous deficient animals. No C2 protein was detected in media from three of the four homozygous deficient animals, but in one, apparent C2 fragments were present. In contrast, intracellular C2 protein was identified in all four homozygous deficient cell cultures. Its mobility on SDS-PAGE was slightly faster than normal. Much less abnormal intracellular C2 protein was recovered from homozygous deficient macrophage monolayers than intracellular C2 protein from normal macrophage monolayers. Monolayers from heterozygous animals produced functional and immunochemical C2 at approximately 30% of the normal rate. Normal rates of biosynthesis and secretion of two other MHC-linked class III antigens, C4 and factor B, were detected in macrophage cultures from homozygous and heterozygous deficient animals. These data suggest that a specific defect, i.e. a structural abnormality in C2 protein, underlies C2 deficiency in guinea pigs.
Evidence for placing the Neu-1 locus within the mouse H-2 complex.
Authors: Figueroa F , Klein D , Tewarson S , Klein J
Published: December 2022

Congenic lines B10.KPA42, B10.KPA132, B10.SNA57, B10.DRB62, and B10.WOA105 carry H-2 haplotypes of wild mice on the genetic background of the strain C57BL/10Sn. Two of the lines (B10.DRB62 and B10.WOA105) have H-2 haplotypes indistinguishable from H-2v of B10.SM. The H-2 haplotype of one line (B10.SNA57) seems to have arisen from H-2v by recombination between the D and Qa-2 loci. The H-2 haplotypes of the remaining two lines probably arose from H-2v by recombination between the C4 and D loci. Since all six and no other lines carry the rare Neu-1a allele, the neuraminidase-1 locus is probably located proximal to the H-2D locus. Typing of H-2s recombinants for the enzyme acid phosphatase liver, the processing of which is controlled by the Neu-1 locus, suggests that the locus resides between the E alpha and D loci, that is in the S region.
Drug transfer into lymphoblasts mediated by liposomes bound to distinct sites on H-2 encoded I-A, I-E, and K molecules.
Authors: Machy P , Pierres M , Barbet J , Leserman L D
Published: December 2022

Protein A from S. aureus was covalently coupled to small sonicated liposomes containing methotrexate (MTX) and carboxyfluorescein (CF). These liposomes were incubated with mitogen-stimulated CBA T or B cells preincubated with one of a panel of murine IgG2 monoclonal antibodies directed at distinct sites on H-2Kk, I-Ak, or I-Ek molecules. Liposomes became bound to the cells as a consequence of the interaction of protein A on the liposome and antibody on the cell. The number of bound liposomes was measured by fluorescence, and the endocytosis of liposomes was evaluated by the MTX-mediated reduction in radiolabeled deoxyuridine incorporation. T cells were sensitive to MTX in liposomes bound to all anti-H-2Kk antibodies tested, but B cells were essentially insensitive, even though they expressed more H-2Kk, bound more liposomes directed at this molecule, and were equally sensitive to free MTX. By contrast, B cells were sensitive to liposome-encapsulated MTX when liposomes were bound to any antibody directed at I-Ak. The situation was more complex with respect to I-Ek. Antibodies directed at one part of the molecule were quite effective at drug delivery, while antibodies directed at other, spatially separate domains were much less effective, even though the number of liposomes bound to these various determinants was the same. The results suggest differential regulation of the internalization of H-2K by T and B cells, and are compatible with the existence of functionally distinct I-E molecules.
Phagocytosis by rat peritoneal mast cells: independence of IgG Fc-mediated and C3-mediated signals.
Authors: Otani I , Conrad D H , Carlo J R , Segal D M , Ruddy S
Published: December 2022

Phagocytosis of sheep erythrocytes (E) bearing rabbit immunoglobulin and rat C3 by rat peritoneal mast cells was quantitated by using 51Cr-labeled E and was confirmed by electron microscopy. The relative importance of IgG Fc receptor interaction in C3-mediated phagocytosis was assessed. Removal of any traces of IgG antibody by absorption of IgM antibody with protein A-Sepharose and absorption of the other reagents with sheep E had no effect on the phagocytosis of C3b- and C3bi-coated cells. IgG antibody enhanced the phagocytosis of intermediates prepared with IgM, purified components and rat C3 (EaIgMClgp4hu oxy2hu3brat) with a graded dose response, but was only additive and not synergistic with C3. The independence of the C3- and Fc-mediated signals was confirmed by using chemically produced polymers of rabbit IgG to inhibit phagocytosis. These polymers, especially tetramers and higher aggregates, completely blocked ingestion of EAIgG, but not that of EAIgMC1423b or -3bi. When IgG was substituted for IgM in the C3b intermediate, the IgG polymers inhibited about 50% of the phagocytosis. Cumulatively, the data demonstrated that in the case of rat mast cells, the stimuli to phagocytosis induced by C3 and IgG are independent; either is sufficient by itself.
Characterization of the rat mast cell beta-adrenergic receptor in resting and stimulated cells by radioligand binding.
Authors: Marquardt D L , Wasserman S I
Published: December 2022

Rat serosal mast cell beta-adrenergic receptors were characterized both functionally by assessing changes in histamine release and cyclic 3',5' adenosine monophosphate (cAMP) levels and directly by radioligand binding studies using [3H]dihydroalprenolol ([3H]DHA), a beta-adrenergic antagonist. Mast cells were obtained by lavage of the pleural and peritoneal cavities of Sprague-Dawley rats and were purified on metrizamide gradients to greater than 95% purity. Resting mast cells stimulated with beta-adrenergic agonists demonstrate a marked rise in cAMP levels after a 15-sec incubation. However, the same concentrations of these agonists have no effect on IgE-mediated mast cell histamine release. [3H]DHA binding to intact mast cells is rapid, reversible, saturable, and stereoselective. The cells possess 40,000 +/- 14,000 beta-adrenergic receptors/cell and demonstrate a binding affinity of 1.58 +/- 0.56 nM for [3H]DHA. Competition studies reveal that 83.5% of the receptors are of the beta 2 subtype and 16.5% are beta 1. Neither sensitization with anti-DNP-BSA IgE nor subsequent challenge with specific antigen alters mast cell beta-adrenergic receptor characteristics. Rat mast cells possesses large numbers of high affinity beta-adrenergic receptors, primarily of the beta 2 subtype, coupled to adenylate cyclase, but the role of these receptors in mast cell secretory events is not yet established.
Subclass restriction of anti-Sm antibodies in MRL mice.
Authors: Eisenberg R A , Winfield J B , Cohen P L
Published: December 2022

The subclass distribution of anti-Sm antibodies in the serum of MRL/Mp-Ipr/Ipr and MRL/Mp- +/+ mice was investigated with a sensitive ELISA technique. In both strains, but particularly in the Ipr mice, anti-Sm antibodies were predominantly of the IgG2a isotype. This preponderance was not an artifact of the sensitivity or specificity of the subclass-specific anti-Sm ELISA, nor did it reflect the subclass distribution of total serum IgG. A plaque-forming cell assay for anti-Sm antibodies also showed a predominance of the IgG2a isotype, indicating that the serum findings could not be explained by differential catabolism of IgG subclasses. Finally, antibodies to double-stranded DNA, as detected by the Crithidia luciliae assay, did not show a restricted subclass distribution. The isotype expression of anti-Sm antibodies in MRL mice must reflect in vivo mechanisms regulating the production of these autoantibodies. The IgG2a restriction in particular suggests an important in vivo role for T cells.
Immunopathology of BCG infection in genetically resistant and susceptible mouse strains.
Authors: Pelletier M , Forget A , Bourassa D , Gros P , Skamene E
Published: December 2022

Natural resistance to Mycobacterium bovis (BCG) is under the control of a single gene, designated Bcg. Resistant (Bcgr) mice prevent multiplication of an i.v. injected inoculum of congruent to 10(4) dispersed BCG cells, whereas progressive multiplication of this pathogen occurs in the first 3 wk of infection in spleens and livers of susceptible (Bcgs) mice. Striking differences in the development of cellular immunity, as measured by granuloma formation in the liver and spleen, delayed-typed hypersensitivity, and a resistance to the challenge with homologous (BCG) and heterologous (Listeria monocytogenes) pathogens, were detected between Bcgr (C3H/HeN and A/J) and Bcgs (C57BL/6J and B10.A) strains. Cellular immune reactions progressively developed in the Bcgs mice, as a response to the increasing bacterial load, whereas greatly inferior levels of acquired immunity were observed in Bcgr strains. These findings support the concept that mice genetically resistant to BCG infection are able to prevent bacterial multiplication without the need for a cellular immune response, whereas genetically susceptible mice will eventually control bacterial multiplication with the acquisition of cellular immunity.
Complement-mediated leukocyte adherence to infective larvae of Dipetalonema viteae (Filarioidea): requirement for eosinophils or eosinophil products in effecting macrophage adherence.
Authors: Haque A , Ouaissi A , Santoro F , des Moutis I , Capron A
Published: December 2022

The present study reports the existence of C-mediated adherence of eosinophils and/or macrophages to filarial infective larvae of Dipetalonema viteae. C3 molecules are present on the surface of the parasite, as shown by immunofluorescence studies. Samples of fNRS depleted of AP of complement by treatment with Zymosan A or of factor B by heating at 50 degrees C for 20 min fail to mediate cell adherence to the parasite. In contrast, fNRS inactivated for CP of complement by the chelating agent EGTA retains its activity in mediating cell adherence to the parasite. There is a significant consumption of factor B and AP of complement when infective larvae are incubated in fNHS. Consumption of C4 of the CP of complement is much lower in the same test. The adherence of macrophages cannot occur without the simultaneous presence of eosinophils, whereas eosinophils probably act alone and not in concert with macrophages. The eosinophil adherence is associated with degranulation. The damage is notably enhanced by replacing the spent eosinophil population with a newly obtained population. In the present test system, mast cells did not adhere to filarial larvae even when mast cell-rich populations were used, nor did they effect macrophage adherence when presented in association with the latter. When eosinophil-enriched cell populations containing less than 1% mast cells were used, cell adherence to filarial larvae still occurred, but the presence of 30% mast cells in such cell populations markedly increased both the rate and level of adherence. We suggest that a cell-mediated adherence and destruction dependent upon the activation of complement via AP, without a requirement for specific antibody, may represent a natural mechanism of parasite killing in a nonimmune host.
Allergic Reactions to Streptococcus Antigens
Authors: Mackenzie George M. , Hanger Franklin M.
Published: December 2022

In a preliminary communication (1) we reported briefly on what we believe to be allergic skin reactions occurring after intracutaneous injections of derivatives of hemolytic and nonhemolytic streptococci. The present paper describes these observations in more detail, and reports extensions of this work carried on during the past two years.

Antigens. Most of the observations were made with derivatives of hemolytic and non-hemolytic streptococci obtained from a variety of sources. A few strains of Staphylococcus aureus and albus, Pfeiffer bacillus, B. coli, B. typhosus, B. paratyphosus A and B were also tested. Apparently the same type of reaction occurs in certain individuals with derivations of some strains of these organisms; some of these filtrates, however, caused reactions which seem to be due to primarily toxic substances, rather than, as in the case of the streptococcus antigens, acquired reactions to substances inactive in non-allergic individuals.
Reactions of the Urinary Bladder in Rabbit Anaphylaxis
Authors: Manwaring W. H. , Marino H. D.
Published: December 2022

Summary and Conclusions

On intravenous injection of specific foreign protein into hypersensitive guinea pigs, there is invariably a sharp and very marked contraction of the urinary bladder, usually sufficient to increase the intracystic pressure about 40 mm. Hg. Relaxation usually sets in during the third minute, but in fatal anaphylaxis the intracystic pressure rarely falls below 20 mm. Hg before the death of the animal.

This marked, prolonged urinary bladder contraction is consistent with the generally accepted view that smooth muscle contractions play a dominant rôle in the production of the anaphylactic syndrome in guinea pigs.

On intravenous injection of specific foreign protein into hypersensitive rabbits, there is, in contrast, in the great majority of cases no recordable increase in the tone of the urinary bladder, even in the rapidly fatal type of anaphylaxis leading to death of the animal in ten to fifteen minutes. In fact, about a third of the rabbits show the opposite phenomenon, a partial or even complete loss of cystic tone.

In about 20 per cent of the rabbits thus far tested, however, a slight suggestion of a cystic contraction was recorded, usually sufficient to increase the intracystic pressure about 1 or 2 mm. Hg. Full relaxation usually took place by the end of the fourth minute.

This uniformly negative or practically negative result is inconsistent with the suggestion that smooth muscle contractions play an appreciable rôle in the production of anaphylactic symptoms in rabbits.
Relation of Anaphylaxis to Immunity
Authors: Manwaring W. H. , Marino H. D. , McCleave T. C. , Boone T. H.
Published: December 2022

Summary and Conclusions

Urinary bladders of goat serum immune dogs, therefore, show no suggestion of an acquired insusceptibility to the histamine-like substances explosively formed or liberated by the hypersensitive liver during the initial stages of horse serum shock. Horse serum immune bladders show no suggestion of an acquired insusceptibility to the hepatic anaphylatoxins of goat serum shock.

This apparent specificity of the acquired fixed-tissue antianaphylatoxic insusceptibility, is in harmony with our conception that the hepatic anaphylatoxin is a specific secondary antigen, presumably of protein nature, conceivably a denaturization product of the primary protein used in sensitization or immunization.
The Relatively Heat Stable Components of Complement
Authors: Whitehead Hugh Robinson , Gordon John , Wormall Arthur
Published: December 2022

Small quantities of ammonia or amines inactivate complement by destroying a relatively heat stable component, not identical with the third component. A fourth component of complement is therefore postulated. The action of ammonia is quite specific, for other alkalies have no similar effect. The possible significance of this new component in explaining difficulties and discrepancies in previous work on complement is discussed. The relatively heat-stable components of serum are much more stable towards ultra-violet light than the heat-labile components.
The Action of Pancreatic Extracts on Complement
Authors: Wormall Arthur , Whitehead Hugh Robinson , Gordon John
Published: December 2022

The proteoclastic and anticomplementary powers of three pancreatic extracts have been compared. The ratio is almost identical for each function. Thus the observation of Grove that the proteoclastic and anticomplementary functions of a pancreatic extract reside in two different substances cannot be confirmed.
Activation requirements of cloned inducer T cells. II. The failure of some clones to respond to antigen presented by activated B cells.
Authors: Dekruyff R H , Cantor H , Dorf M E
Published: December 2022

Inducer/helper T cells recognize nominal antigen in association with Ia on the surface of the antigen-presenting cell (APC). Recent studies have shown that B cells can effectively function as APC. In the present study we have assessed the ability of cloned inducer T cells to discriminate between activated B cells or splenic macrophages as APC. We found that most of the clones tested demonstrated an equivalent response to antigen presented by activated B cells or splenic adherent cells. Some clones were very efficiently stimulated by antigen presented by activated B cells, whereas other clones failed to respond or responded very poorly when activated B cells were used to present antigen. We attempted to determine the mechanism responsible for the inability of certain clones to proliferate in response to antigen presented by activated B cells.
Oligopeptide antigens of the angiotensin lineage compete for presentation by paraformaldehyde-treated accessory cells to T cells.
Authors: Buus S , Werdelin O
Published: December 2022

The heptapeptide antigen angiotensin III can be presented to guinea pig T cells by paraformaldehyde-treated antigen-presenting cells, which are incapable of processing antigens and presumably cannot even ingest them. We demonstrate here that the decapeptide angiotensin I can outcompete angiotensin III for presentation by paraformaldehyde-treated antigen-presenting cells. It seems likely that the competition is for a site on the surface of the presenting cell. This extends earlier findings of competition for presentation between antigens. We also demonstrate that the antigens of the angiotensin series are highly susceptible to proteolytic destruction in cultures containing prefixed accessory cells. The proteases responsible for the destruction of these peptides are apparently located in the plasma membrane of accessory cells. These enzymes represent a methodologic problem in studies of competition between antigens for presentation; but since they presumably are active also in untreated cells, they may play a physiologic role in the normal immune response.
Identification of an antigenic determinant within the phylogenetically conserved triprolyl region of myelin basic protein.
Authors: Potter N T , Hashim G A , Day E D
Published: December 2022

Synthetic peptide SP6 (RTPPPSG), comprising amino acid residues 98-103-Gly of myelin basic protein (MBP), and a series of peptide analogs were used to probe the structural requirements for antigenicity of a highly conserved region of a self protein. By means of a liquid-phase radioimmunoassay, antibody responses directed toward this determinant in both multi-specific anti-MBP and monospecific anti-peptide antisera were measured. The specificities of the antibodies present in the anti-MBP and anti-peptide antisera were examined by an equilibrium competitive inhibition radioimmunoassay by using the set of related peptides, as well as intact MBP from different species. Although the fine specificities of the reagent antisera differed, competitive inhibition analyses with intact MBP revealed a cross-reactive determinant involving residues 99-100 (Thr-Pro). This suggests that the neighborhood of the triprolyl region of MBP, despite its strong phylogenetic conservation, serves as an immunogen for humoral responses whether presented as a hapten-carrier conjugate or in the context of intact MBP. The latter supports the contention that the general antigenicity of a protein need not require sequence differences between the immunizing protein and its counterpart in the host.
Spontaneous interstitial nephritis in kdkd mice. II. Characterization of a tubular antigen-specific, H-2K-restricted Lyt-2+ effector T cell that mediates destructive tubulointerstitial injury.
Authors: Kelly C J , Korngold R , Mann R , Clayman M , Haverty T , Neilson E G
Published: December 2022

In this report, we have examined the effector T cell repertoire in the spontaneous interstitial nephritis of kdkd mice. Lymph node cells from nephritic kdkd mice are capable of transferring this disease into thymectomized, irradiated, and bone marrow-reconstituted CBA/Ca recipients. CBA/Ca mice do not spontaneously develop interstitial nephritis and are normally resistant to the adoptive transfer of nephritic cells, a resistance that in the short term can be attenuated with low-dose cyclophosphamide. We therefore used delayed-type hypersensitivity responses and direct transfer of immune cells under the renal capsule to characterize nephritogenic effector cells from kdkd donor mice. Lyt-2+, L3T4- T cells from the peripheral lymphoid organs of nephritic kdkd mice, after adoptive transfer into cyclophosphamide-pretreated CBA/Ca recipients, mediate an antigen-specific delayed-type hypersensitivity response to renal tubular basement membrane antigens. These cells are restricted by gene products in H-2Kk; they are also present in nephritic, but not in control kidneys. We have also observed this same phenotypic subpopulation of kdkd lymphocytes mediate a destructive interstitial renal lesion within 7 days of being placed under the kidney capsule of CBA/Ca mice. These findings suggest that T lymphocytes reactive to a parenchymal tubular antigen are of substantial importance in the development of spontaneous interstitial nephritis in kdkd mice.
Effects of bacterial lipopolysaccharide on the binding of lymphocytes to endothelial cell monolayers.
Authors: Yu C L , Haskard D , Cavender D , Ziff M
Published: December 2022

Preincubation of human umbilical vein endothelial cell (EC) monolayers with 1 ng to 10 micrograms/ml lipopolysaccharide (LPS) increased the binding of T lymphocytes to EC. The effect was maximal at LPS concentrations of 0.1 to 10 micrograms/ml, and occurred with LPS derived from Escherichia coli (serotypes 0111:B4 and 0127:B8), Shigella flexneri (serotype 2a), Serratia marcescens (serotype 0:3), and Yersinia entercolitica (serotype 0:3). The increased binding appeared to be mediated primarily through an action on EC; preincubation of T cells rather than EC with LPS did not lead to enhanced binding. The onset of enhanced binding was very rapid, being observed after 2 to 3 min of preincubation and becoming maximal after 1 hr. EC were unresponsive to LPS after fixation with 2% paraformaldehyde-L-lysine-periodate and also when the LPS was incubated with EC at 4 degrees C. Enhanced binding was seen with lipid A and with LPS from Salmonella minnesota Re 595 (mainly lipid A) and was abolished by conjugation with polymyxin B. The observed increase in the binding of lymphocytes to EC exposed to LPS suggests that the lymphocytopenia induced by endotoxemia may result from augmentation of the adherence of lymphocytes to altered endothelium.
Hypervariable region peptides variably induce specific anti-idiotypic antibodies: an approach to determining antigenic dominance.
Authors: Seiden M V , Heuckeroth R , Clevinger B , McMillan S , Lerner R , Davie J M
Published: December 2022

Rats and rabbits were immunized with synthetic peptides corresponding to the VH hypervariable regions of several alpha (1----3) dextran-specific antibodies from mice to study the efficacy of synthetic peptides in the generation of site-specific anti-idiotypic reagents. Synthetic peptides were made which corresponded to the HV1, HV2, and HV3 hypervariable regions of the heavy chain of M104 (IdX+, IdI-(M104)+), HV2 of HDex 14 (IdX-), and HV3 of J558 (IdX+, IdI(J558)+). The HV1(M104) peptide sequence is found in all dextran-specific immunoglobulins examined and the HV2 and the HV3 peptides span the regions implicated in IdX and IdI expression, respectively. Sera from many rabbits and rats indicate that all five peptides are immunogenic. Antisera to HV3 peptides show excellent binding to the appropriate myeloma proteins, with antisera to the HV3(M104) peptide demonstrating little binding to proteins that differ in HV3 sequence. Antisera generated against HV1(M104) and both HV2 peptides show weak cross-reaction to the appropriate proteins; however, these sera are not idiotypic because they cross-react with immunoglobulins with very limited sequence homology. Thus, it appears that some, but not all synthetic peptides from hypervariable regions will be capable of generating antisera with useful anti-idiotypic specificities. This may reflect differences in the intrinsic antigenicity of various parts of the VH region.
Neutrophil-mediated solubilization of the subendothelial matrix: oxidative and nonoxidative mechanisms of proteolysis used by normal and chronic granulomatous disease phagocytes.
Authors: Weiss S J , Curnutte J T , Regiani S
Published: December 2022

Both normal and chronic granulomatous disease (CGD) neutrophils were able to degrade the subendothelial matrix secreted by human endothelial cells via an elastase-dependent process. In the absence of the plasma antiproteinase, alpha-1-proteinase inhibitor (alpha-1-PI), normal neutrophils protect their released elastase from inactivation by using the chlorinated oxidants hypochlorous acid and endogenous N-chloroamines to suppress the antiproteinase's activity. In contrast, CGD neutrophils were unable to generate either class of chlorinated oxidant or to inactivate the porcine pancreatic elastase inhibitory capacity of alpha-1-PI unless the cells were supplemented with exogenous hydrogen peroxide. Despite the reliance of normal neutrophils on chlorinated oxidants to inactivate alpha-1-PI, neutrophils triggered in the presence of agents that block the generation of these reactive species continued to degrade the subendothelial matrix at a suppressed but significant rate in the presence of a 50-fold excess of the antiproteinase. The continued solubilization of the matrix by normal neutrophils was not due to the incomplete inhibition of oxidant generation because triggered CGD neutrophils were also able to degrade the matrix in the presence of excess alpha-1-PI. If CGD neutrophils were stimulated in the presence of an exogenous source of H2O2 and alpha-1-PI, the proteolytic potential of the cells was identical to that observed with normal stimulated neutrophils. We conclude that normal neutrophils can enhance their ability to degrade the subendothelial matrix by oxidatively protecting elastase from inactivation by alpha-1-PI but both normal and CGD neutrophils possess non-oxidatively linked mechanisms for sequestering and using elastase to mediate proteolytic effects in the presence of native antiproteinase.
Macrophage-mediated fungistasis in vitro: requirements for intracellular and extracellular cytotoxicity.
Authors: Granger D L , Perfect J R , Durack D T
Published: December 2022

Macrophage cytotoxicity for Cryptococcus neoformans was investigated by culturing mouse peritoneal macrophages with a thin-capsuled clone of cryptococcus under conditions permitting efficient phagocytosis. Yeast replication was quantitated by electronic particle counting after detergent lysis of macrophages, and viability was determined by quantitative plate counts. Under appropriate conditions, reproduction was completely inhibited; stasis began at 2 hr after addition of yeasts and lasted for 30 hr. During this time organisms in medium alone proliferated rapidly, doubling their number every 2.5 hr. After removal from macrophages, 60 to 100% of macrophage-inhibited cryptococci formed colonies, indicating that the cytotoxic effect was primarily fungistatic. When yeast cells were removed from macrophages, replication recommenced within 5 hr. Supernatant medium from fungistatic co-cultures was not inhibitory for fresh yeast cells. Conditions required for complete fungistasis were 1) peritoneal macrophages induced by peptone from BCG-infected mice, 2) endotoxin in nanogram per milliliter range added to serum-containing cell culture medium, 3) confluent macrophage monolayers, and 4) macrophage:cryptococci ratios of 20 to 100:1. Fungistasis occurred without phagocytosis but was more efficient when cryptococci were engulfed. For efficient fungistasis, macrophages must differentiate to and be maintained in the activated state. These results with yeast cells agree with the known requirements for macrophage effector function against neoplastic target cells.
Immunosuppressive effects of glycosylation inhibiting factor on the IgE and IgG antibody response.
Authors: Akasaki M , Jardieu P , Ishizaka K
Published: December 2022

Glycosylation inhibiting factor (GIF) was purified from culture filtrates of a T cell hybridoma, 23A4, by affinity chromatography on anti-lipomodulin Sepharose. The factor exhibited phospholipase inhibitory activity upon dephosphorylation. Immunization of BDF1 mice with aluminum hydroxide gel (alum)-absorbed dinitrophenyl derivatives of ovalbumin (DNP-OA) resulted in persistent IgE and IgG antibody formation. However, repeated injections of the affinity-purified GIF into the DNP-OA-primed mice beginning on the day of priming prevented the primary anti-hapten antibody responses of both the IgE and the IgG1 isotypes. Treatment with GIF also diminished on-going IgE antibody formation in the DNP-OA-primed mice. The treatment changed the nature of IgE-binding factors formed by BDF1 spleen cells. Incubation of spleen cells from OA + alum-primed mice with OA resulted in the formation of IgE-potentiating factor, whereas spleen cells of OA-primed, GIF-treated mice formed IgE-suppressive factor upon antigenic stimulation. It was also found that Lyt-2+ T cells in the OA-primed, GIF-treated mouse spleen cells released GIF, which had affinity for OA and bore I-Jb determinant(s). Transfer of a Lyt-1+ cell-depleted fraction of the OA-primed, GIF-treated mouse spleen cells into naive syngeneic animals resulted in suppression of the primary anti-DNP IgE antibody response of the recipients to alum-absorbed DNP-OA, but failed to affect the anti-DNP antibody response to DNP-keyhole limpet hemocyanin. The results indicate that GIF treatment during the primary response to OA facilitated the generation of antigen-specific suppressor T cells.
Wheat germ agglutinin activates human T lymphocytes by stimulation of phosphoinositide hydrolysis.
Authors: Clevers H C , de Bresser A , Kleinveld H , Gmelig-Meyling F H , Ballieux R E
Published: December 2022

Recently, it has become evident that stimulated phosphoinositide (PI) hydrolysis plays a crucial role in early T lymphocyte activation. We have investigated the effects of the nonmitogenic lectin wheat germ agglutinin (WGA) on several parameters associated with PI hydrolysis in human T cells. It was found that WGA was as effective as anti-T3 antibody and PHA in producing a rise in cytosolic free Ca++ ((Ca++)i) in blood T cells and in cells of the T cell line CCRF-CEM. It was inferred that identical cells within the blood T cell preparation responded to each of the three agents, refuting the contention that WGA only stimulated a subfraction of circulating mature T lymphocytes. WGA-induced, but not PHA-induced rises in (Ca++)i could be blocked completely by N-acetyl-D-glucosamine, demonstrating that the sugar-binding characteristics of the lectin dictate its action on T lymphocytes. Anti-T3 antibody, PHA, and WGA all initiated inositol phosphate formation in blood T cells, indicating that each of the agents stimulated PI hydrolysis. The combination of WGA with nonmitogenic amounts of phorbol-12-myristate-13-acetate resulted in strong mitogenicity. It is concluded that WGA, like anti-T3 antibody and PHA, is a pan-T activator of PI hydrolysis.
Role of the LFA-1 molecule in cellular interactions required for antibody production in humans.
Authors: Fischer A , Durandy A , Sterkers G , Griscelli C
Published: December 2022

The lymphocyte function-associated antigen 1 (LFA-1) has been shown to play a role in various T cell functions in mice and humans including cytotoxicity, and proliferation to allogeneic cells and foreign antigens. These functions have been defined with specific monoclonal antibodies and were additionally confirmed by the investigation of patients with inherited deficiency in membrane LFA-1 expression. In this paper, we report our studies on the potential role of the LFA-1 molecule in T lymphocyte-dependent antibody responses. In a patient with a complete lack of membrane expression of LFA-1, there was no in vivo antibody response to vaccinal antigens such as tetanus, diphtheria toxoids, and polio virus, and no in vivo or in vitro antibody production to influenza virus, whereas serum immunoglobulin levels and antibodies to polysaccharides (isohemagglutinins, antibody to mannan, and a polysaccharide from Candida albicans) were detected in correlation with in vitro production of anti-mannan antibody. The defective antibody response to polypeptides was not secondary to poor antigen-specific T proliferation, because the latter was found to be present. Similarly, in vitro antibody production to influenza virus of normal cells was blocked by several anti LFA-1 monoclonal antibodies specific for the alpha subunit of the molecule, if they were added from the beginning of the culture. The antibody production blockade could be achieved with monoclonal antibody concentrations that partially preserved T cell proliferation. The helper effect of an influenza virus-specific helper T cell clone was also blocked. The targets of the blockade were shown by incubation experiments to be T cells and monocytes. In contrast, anti-LFA-1 monoclonal antibodies had no effect on pokeweed mitogen-induced B cell maturation into immunoglobulin-containing cells and on the anti-mannan antibody production. These combined data demonstrate that the LFA-1 molecule plays a role in T cell dependent antibody production to polypeptidic antigens but not in the antibody response to polysaccharides, although the antibody response to mannan is T cell dependent. It is proposed that the LFA-1 molecule is required to some extent for a antigen-presenting cells-T lymphocyte interaction and for the maintenance of a close association between antigen-specific helper T cells and small resting B lymphocytes. Polysaccharidic antigens that exhibit repetitive antigenic determinants might cross-link membrane immunoglobulins on B lymphocytes, thus allowing B cells to pass through a first step of activation requiring cognate T-B cell interaction.
Isolation and analysis of primitive hemopoietic progenitor cells on the basis of differential expression of Qa-m7 antigen.
Authors: Bertoncello I , Bartelmez S H , Bradley T R , Stanley E R , Harris R A , Sandrin M S , Kriegler A B , McNiece I K , Hunter S D , Hodgson G S
Published: December 2022

In the presence of the hemopoietic growth factor CSF-1, the later committed cells of the macrophage lineage can be detected by their ability to form small colonies in clonal agar culture (CFCCSF-1). Synergistic factors have been described that in combination with CSF-1 stimulate developmentally early hemopoietic progenitor cells of high proliferative potential (HPP-CFC). By using a monoclonal antibody to the Qa-m7 antigenic determinant, we investigated and compared the expression of Qa-m7 on CFCCSF-1 and on HPP-CFC of two types that grow in response to either 1) CSF-1 plus synergistic factor from human placenta-conditioned medium (HPP-CFCHplac+CSF-1) or 2) CSF-1 plus synergistic factor from conditioned medium of the WEHI-3 myelomonocytic cell line (HPP-CFCW+CSF-1). We have shown that HPP-CFC of both types express relatively more Qa-m7 antigen than CFCCSF-1 and can be separated and enriched on this basis by discontinuous buoyant density centrifugation and fluorescence-activated cell sorting of normal bone marrow. Significant enrichments of HPP-CFCHPlac+CSF-1 (43.5-fold) and HPP-CFCW+CSF-1 (28.8-fold) have been achieved with cloning efficiencies of HPP-CFC in the most enriched fractions reaching 4 to 5%. These results clearly illustrate the fact that there are populations of progenitor cells from normal, unperturbed bone marrow that strictly require a combination of two hemopoietic growth factors (CSF-1 plus synergistic factor) in order to be detected.
Immunoregulatory molecules and IL 2 receptors identified in multiple sclerosis brain.
Authors: Hofman F M , von Hanwehr R I , Dinarello C A , Mizel S B , Hinton D , Merrill J E
Published: December 2022

Frozen brain specimens from eight multiple sclerosis (MS) patients were examined for the presence of leukocytes and their cell products by using a panel of monoclonal antibodies. The results presented here demonstrated that in the MS lesion, there was a marked accumulation of HLA-Dr-positive (Ia-positive) cells. These Ia-positive cells were identified as being glial fibrillary acidic protein positive by using double staining methods. Furthermore, the cells in the MS lesion expressed the interleukin 2 (IL 2) receptor, as identified by the anti-TAC monoclonal antibody. The cells in the region of the plaque also exhibited positive staining with antibodies to IL 1, IL 2, and prostaglandin E. Neither normal brain nor brain specimens from progressive multifocal leukoencephalopathy or Alzheimer's disease showed such patterns of staining. These results suggest that stimulated cells are present in the MS brain, thus implicating an active immune mechanism in the pathogenesis of MS.
Interleukin 1 is present in normal human epidermis.
Authors: Hauser C , Saurat J H , Schmitt A , Jaunin F , Dayer J M
Published: December 2022

We investigated the presence of interleukin 1 (IL 1)-like molecules in normal unstimulated human epidermal tissue. Epidermis from 21 healthy individuals that was prepared by two different methods showed prostaglandin E2 (PGE2) and collagenase stimulating activity for human dermal fibroblasts. All epidermal extracts tested were positive for thymocyte comitogenic activity (lymphocyte activating factor; LAF). Removal of the horny layer decreased epidermal IL 1-like activity. In contrast to epidermal tissue, freshly isolated peripheral blood mononuclear cells (PBMC) contained no detectable PGE2 stimulatory activity. They could, however, produce PGE2 stimulatory activity after culture and stimulation with phytohemagglutinin (PHA) and concanavalin A (Con A). Little membranous IL 1-like activity could be detected in epidermal extracts when using a method that has previously rendered membranous IL 1 from murine proteose peptone-elicited peritoneal macrophages. Gel filtration chromatography yielded double peaks at m.w. approximately 30,000 and approximately 17,000 for all three activities. High pressure liquid chromatography (HPLC) analysis identified two species with a m.w. of approximately 17,000, and one approximately 30,000 species nondissociable in detergent, all having superposable PGE2 and collagenase stimulatory as well as LAF activity. These results establish the existence of IL 1-like molecules, together with a possible precursor, in normal human epidermis. The release of these preformed epidermal IL 1 stores might be important in vivo.
Structural analysis of human complement protein H: homology with C4b binding protein, beta 2-glycoprotein I, and the Ba fragment of B2.
Authors: Kristensen T , Wetsel R A , Tack B F
Published: December 2022

We report here a partial primary structure for human complement protein H. Tryptic peptides comprising 27% of the H molecule were isolated by conventional techniques and were sequenced (333 amino acid residues). Several mixed-sequence oligonucleotide probes were constructed, based on the peptide sequence data, and were used to screen a human liver cDNA library. The largest recombinant plasmid (pH1050), which hybridized with two probes, was further characterized. The cDNA insert of this plasmid contained coding sequence (672 bp) for 224 amino acids of H. The 3' end of this clone had a polyadenylated tail preceded by a polyadenylation recognition site (ATTAAA) and a 3'-untranslated region (229 bp). Four regions of internal homology, each about 60 amino acids in length, were observed in the derived protein sequence from this cDNA clone, and a further seven from the tryptic peptide sequences. The consensus sequence for each of the repetitive units of H was four cysteines, two prolines, three glycines, one tryptophan, and two tyrosines/phenylalanines. Based on the mole percent values for each of these amino acids, it is likely that H is composed of about 20 repetitive units of this nature. Furthermore, the repetitive unit of H shows pronounced homology with the Ba fragment of B, the C4b binding protein, and beta 2-glycoprotein I. Therefore, it seems that at least portions of these proteins have evolved from a common ancestral DNA element.
The nucleotide sequence and comparative analysis of the H-2Dp class I H-2 gene.
Authors: Schepart B S , Takahashi H , Cozad K M , Murray R , Ozato K , Appella E , Frelinger J A
Published: December 2022

We present the complete nucleotide sequence and the deduced amino acid sequence of the H-2Dp class I gene. This gene, which was cloned from a B10.P genomic DNA library, encodes and intact, functional H-2Dp molecule. Comparative analysis of the Dp sequence with other class I sequences reveals both similarities and differences. This analysis also shows that these genes exhibit D region-specific, locus-specific, as well as allele-specific sequences. The H-2Dp nucleotide sequence is greater than 90% homologous to the H-2Ld and H-2Db genes and only approximately 85% homologous to the H-2Dd gene. The K region and Qa region genes are less homologous. The 3' noncoding sequences appear to be region-specific. All of the previously described D region genes, Db, Ld, and Dd, possess the B2-SINE Alu-like repetitive sequence, as does Dp. Thus, this B2 repeat is a region-specific marker present in all D region genes studied so far. The additional polyadenylation site found in the H-2Dp gene starting at nucleotide 4671, which is homologous to non-D region sequences, as well as unique protein Dp coding sequences, make this gene an interesting model for studying the evolution of polymorphism and structure/function relationships in the class I gene family.