Selective platelet immune retention.

Inbred rats were immunized with Freund's Adjuvant containing proteins of Mycogacterium tuberculosis, Mycobacterium butyrium, or dinitrochlorobenzene (DNCB). Native arterial blood was then passed over glass beads coated with those antigens. The platelet retention on the glass beads was measured. Rats immunized with Complete Freund's Adjuvant developed accelerated platelet retention on beads coated with protein derivatives of tuberculosis (PPD) after just 18 seconds of blood flow. Rats immunized twice maintained selective retention longer than those immunized once. The test animals developed no cutaneous hypersensitivity nor precipitin lines on Ouchterlong gels against PPD. Rats sensitized to DNCB had accelerated platelet retention on DNCB-coated beads. Results were temperature and complement dependent, and antigen-specific. Heparin caused a dose-dependent inhibition of the accelerated retention. PPD potentiated the ADP-induced aggregation of plateletrich plasma (PRP) from immunized rats. These experiments suggest that platelets react selectively to antigens in the intravascular spaces in immune reactions.     

Liposomes in Immunology: Multilamellar Phosphatidylcholine Liposomes as a Simple, Biodegradable and Harmless Adjuvant Without any Immunogenic Activity of its own

Multilamellar phosphatidylcholine liposomes were coated with the antigens human serum albumin (HSA) or bovine gamma globulin (BGG) simply by suspending the liposomes in a solution of the antigens.

Antigen coated phosphatidylcholine liposomes showed nearly the same adjuvant activity after intravenous injection as liposomes of a more complex phospholipid composition.

Since phosphatidylcholine liposomes are biodegradable, harmless, easily obtainable, have no immunogenic activity of their own and may be administered intravenously, they seem to be a promising immunoadjuvant.     

Mycobacterium vaccae immunization to OVA sensitized pregnant BALB/c mice suppressed placental and postnatal IL-5 and inducing IFN-gamma secretion

Although the development of atopy in the newborn is determined by a multitude of factors, an intense Th1 stimulus early in life could be protective by facilitating a switch away from Th2. Aimed to determine the effect of single Mycobacterium vaccae (M. vaccae) immunization to OVA-sensitized pregnant mice on IL-5 and IFN-γ secretion from placental lymphocytes and splenocytes of offspring.

Pregnant BALB/c mice were divided into 4 groups, OVA-sensitized + M. vaccae immunized, OVA-sensitized, M. vaccae immunized and controls. Sensitization with OVA was initiated before mating, and aerosol OVA challenge were performed during pregnancy. M. vaccae immunization was performed on the 12^th day of pregnancy. IL-5 and IFN-γ levels of placental lymphocytes were analyzed on the 18^th day of pregnancy and splenocytes of offspring on the 2^nd and 28^th days during postnatal period. A single administration of M. vaccae to OVA-sensitized pregnant mice downregulated IL-5 secretion and induced IFN-γ secretion from placental lymphocytes. On the other hand, after M. vaccae immunization downregulation of IL-5 levels and upregulation of IFN-γ secretion persisted in offspring when determined on 2^nd and 28^th days of life. Vaccination with M. Vaccae to OVA-sensitized pregnant BALB/c mice prevented Th2 immune responses by enhancing secretion of IFN-γ and lowering IL-5 levels during pregnancy and the effect persisted during the postnatal period in offspring.     

Induction of interstitial nephritis in rats by basement membrane of human origin

A study was conducted to investigate nephritogenic tubular basement membrane antigens common to human and rat kidneys. Brown Norway (BN) rats were immunized with human renal basement membrane in complete Freund's adjuvant simultaneously with Bordetella pertussis vaccine. The immunized rats developed polyuria and increased levels of serum creatinine one week after the second immunization. Renal histology at this time revealed marked, acute tubulointerstitial nephritis with linear deposition of IgG and C3 along the tubular basement membrane and Bowman's capsule, but not along the glomerular basement membrane. Rats with this tubulointerstitial nephritis rapidly developed antibodies against renal antigens from normal BN rats such as tubular basement membrane and proximal tubule brush border, however antibodies to glomerular basement membrane appeared later. Western blotting using the same rat sera detected a 145-kDa antigen from 8 M urea-solubilized human renal basement membrane and 120-kDa, 135-kDa and 145-kDa antigens from 8 M urea-solubilized BN rat renal basement membrane. This suggests that renal basement membranes of human and rat origin have common antigens involved in the pathogenesis of tubulointerstitial nephritis.     

Book Reviews

Immunological Tolerance, (British Medical Bulletin, Volume 32, No. 2, May, 1976), D. W. Dresser, ed.

Immune Reactivity of Lymphocytes (Development, Expression and Control), M. Feldman, A. Globerson, ed.

Immunogenetics and Immunodeficiency, B. Benacerraf, ed.

Manual of Clinical Immunology, N.R. Rose, H. Friedman, eds.

Immunobiology of the Tumor-Host Relationship, R.T. Smith, M. Landy, eds.

The White Cell, M. L. Cline

Transfer Factor, Basic Properties and Clinical Applications, S. Asbher, A. Gotlieb, Ch. H. Kirkpatrick eds.

The Immune System of Secretions, T.B. Tomasi, Jr.

Surgical Immunology, A.M. Minister, ed.

Comparative Immunology, E.L. Cooper

The Immune System A Course on the Molecular and Cellular Basis of Immunity, M.J. Hobart, I. McConnell, eds.

Immunological Response of the Female Reproductive Tract, B. Cinader, A. de Week

Patch Testing, S. Fregert, H. J. Bandmann, eds.

The Role of Immunological Factors in Infectious, Allergic, and Autoimmune Processes, R. F. Beers, Jr., E. G. Bassett, Eds.     

Cellular and Humoral Hypersensitivity to Adrenal Antigen in Experimental Adrenalitis

A role for specific cellular, as well as humoral immunity has been suggested in experimental adrenalitis. This study was performed to seek a correlation between cellular and humoral immunity in experimental adrenalitis of the guinea pig.

34 guinea pigs (GP) were arranged into 4 experimental groups. One group (11 GP) was immunized with a single injection of 250 mg homologous adrenal antigen (HAA) in complete Freund's adjuvant (CFA). A second group (6 GP) was similarly immunized at 1 and 14 days. A third group (9 GP) received 3 such injections at 1, 14, and 21 days. The fourth group (8 control GP) received RPMI-1640 in CFA. The following were performed on all groups 10 days after the last injection: lymphocyte response to PHA and HAA; HAA-specific macrophage migration inhibition (MIF); antibody titers to HAA by hemagglutination; and histopathology of adrenal, thyroid and testis.

Antibody titers reached a mean level of 500 in each of the 3 HAA-immunized groups. In the single injection group, MIF activity and response to PHA were significantly increased when compared to the other immunized groups and to controls. Histopathologic changes were seen in adrenal glands of all immunized groups, but were most remarkable in the single injection group. Progressively fewer changes were observed in double and triple immunized groups. Antibody titers and histological changes were not found in controls.

Histopathology correlated better with cell-mediated immune parameters than with specific antibody titers; this suggests that cell-mediated mechanisms may be the more important factor in pathologic lesions of experimental adrenalitis.     

Modification of the Immunologic Properties of the Cell Surface

Living CF₁ mouse-transplantable spontaneous mammary adenocarcinoma cells were modified with glutaraldehyde, formalin, 2,4,6-trinitrophenylate, Vibrio cholerae neuraminidase, iodoacetate, heparin, histamine and adenosine 3'5'-monophosphate (cAMP), then used to immunize syngeneic CF₁ mice.

Animals immunized with the fixed (formalinized or glutaraldehyde fixed) neuraminidase-treated cells or the membrane of these cells, rejected two challenging doses of 10⁸ viable unmodified adenocarcinoma cells. Animals immunized with adenocarcinoma cells treated with neuraminidase (170 IU/5×10⁵ cells) or with the spontaneous adenocarcinoma-cell surface glycoprotein, rejected the first challenging dose but developed tumors and died on the second challenge with the viable untreated adenocarcinoma cells. Animals immunized with the adenocarcinoma cells pretreated with trinitrophenylate, glutaraldehyde or formalin, developed temporary resistance to the spontaneous mammary adenocarcinoma.

Adenocarcinoma cells pretreated with NaF, iodoacetate, heparin, EDTA, Colchicine or histamine showed reduced oncogenicity and stronger resistance in mice to the development of a mammary tumor than to a smaller number (10³ AdCa cells) of untreated viable adenocarcinoma cells. Cells treated with adenosine 3'5'-monophosphate accelerated tumor development.     

Induction of Interstitial Nephritis in Rats by Basement Membrane of Human Origin

A study was conducted to investigate nephritogenic tubular basement membrane antigens common to human and rat kidneys. Brown Norway (BN) rats were immunized with human renal basement membrane in complete Freund's adjuvant simultaneously with Bordetella pertussis vaccine. The immunized rats developed polyuria and increased levels of serum creatinine one week after the second immunization. Renal histology at this time revealed marked, acute tubulointerstitial nephritis with linear deposition of IgG and C, along the tubular basement membrane and Bowman's capsule, but not along the glomerular basement membrane. Rats with this tubulointerstitial nephritis rapidly developed antibodies against renal antigens from normal BN rats such as tubular basement membrane and proximal tubule brush border, however antibodies to glomerular basement membrane appeared later. Western blotting using the same rat sera detected a 145 kDa antigen from 8 M ufea solubilized human renal basement membrane and 120-kDa, 135 kDa and 145 kDa antigens from 8M urea solubinzed BN rat renal basement membrane. This suggests that renal basement membranes of human and rat origin have common antigens involved in the pathogenesis of tubulointerstitial nephritis. Acta Pathol Jpn 39: 551 557, 1989.     

Adjuvant and Immunostimulating Activities (in the Absence of Freund's Incomplete Adjuvant) of Chemically Modified Low Molecular Weight Mycobacterial Peptidoglycans

Relatively law molecular wight peptidoglycan fragments extracted from two strains of Mycobacterium tuberculosis var. hominis were chemically coupled with lauric acid. The fatty acid conjugates were compared with the native substances with respect to some immunopotentiating activities.

In vitro, the mitogenic effect on murine spleen lymphocytes was significantly enhanced following conjugation. One of the lauric acid conjugates stirmlated, upon intravenous administration in mice, the formation of antibody-producing cells in the spleen, while the rative substance was devoid of such activity. In adjuvanticity tests performed in the guinea pig in the absence of mineral oil, the fatty acid conjugates generally exerted a higher adjuvant effect on antibody production on delayed trpe hypersensitivity reactions than did the native preparations.     

The Influence of the Shape of Phagocytes on their Adhesiveness

Glucose is known to depress the adhesiveness to glass as well as the phagocytic activity of neutrophils and macrophages. Contact angle measurements indicate that this is not due to any decrease in cell surface free energy. The decreased adhesiveness to glass and depressed phagocytic activity under the influence of glucose are instead found to be due to the fact that glucose causes the phagocytes to retract their pseudopods and to become spherical in shape. It can be shown that a spherical shape tends to enhance the electrostatic repulsion between cells and between cells and other surfaces, while pseudopods help to overcome that electrostatic repulsion.

It is also shown that heparin does not interfere with the normal bizarre shape of neutrophils, while other anticoagulants such as ADP, CPD, and EDTA(for explanation of the symbols, see below) cause the neutrophils to become spherical. This explains why only when heparin is used as an anticoagulant, neutrophils will adhere to glass or nylon surfaces, and thus can be removed from whole blood by filtration through these media.

Finally, it could be demonstrated that platelet adhesiveness is also caused by the induction of a spiculated cell shape, rather than by a change in cell surface free energy.

It was previously noted by one of us (1) that human neutrophils, under in vitro conditions of high (4 and 8mg/ml) glucose levels, manifested a significant depression of their phagocytic activity as well as a decreased adhesiveness to glass surfaces. A similar effect of glucose on guinea pig macrophages has also been observed (2). We have since established that the depressed phagocytic activity and the decreased adhesiveness to glass at high glucose levels are not due to changes in the surface free energy of the phagocytes (see belaw). In view of the important role played by pseudopodic protrusions of the electro-negatively charged cells in their adhesion to other negatively charged surfaces (3), we proceeded to investigate the influence of glucose and a number of other additives on the shape of phagocytic cells.     

Retention of platelets by glass bead filters

Whole blood was passed through glass bead filters and the retention of platelets studied under various experimental conditions. Platelet retention was similar over a wide range of blood flow rates; however, it was decreased at fast rates of flow. Platelet retention was not influenced by the haematocrit value of the blood over a wide range. Retention in filters containing siliconized beads was similar to that in filters with untreated beads. Platelet retention was inhibited by the addition of citrate or adenosine to the blood before passage through the filter and was also less when the blood was cooled. Heparin and phenylindandione therapy did not affect the results.There was a wide range of results of platelet retention tests with normal blood samples and considerable variability between the results of duplicate samples from the same donor. Repeated testing of normal donors over a three-month period gave results which varied widely within the normal range. Twenty-two of 40 uraemic blood samples showed decreased platelet retention. The mechanism of platelet retention from whole blood by glass bead filters is not clear but may be related to platelet aggregation induced by adenosine diphosphate or other aggregating agents.     

Central serotonin 3 receptors play an important role in the modulation of nociceptive neural activity of trigeminal subnucleus caudalis and nocifensive orofacial behavior in rats with persistent temporomandibular joint inflammation

The role of central serotonin 3 receptors on neural activities recorded from superficial laminae of trigeminal subnucleus caudalis/upper cervical spinal cord junction region was investigated using rats with (Complete Freund's Adjuvant day 7 group) or without (non-Complete Freund's Adjuvant group) persistent temporomandibular joint inflammation evoked by Complete Freund's Adjuvant for 7 days. We identified two types of units, Deep-wide dynamic range units and Skin-wide dynamic range units from extracellular recordings. Deep-wide dynamic range units have mechanoreceptive fields in the deep craniofacial tissues including masseter muscle but do not have cutaneous mechanoreceptive fields. Deep-wide dynamic range unit discharges evoked by the formalin injection into masseter muscle were significantly enhanced in the late phase in Complete Freund's Adjuvant day 7 group. Discharges of Skin-wide dynamic range units evoked by the noxious pinch stimulation to facial skin in Complete Freund's Adjuvant day 7 group were significantly enhanced compared with those in non-Complete Freund's Adjuvant group. Topical administration of central serotonin 3 receptor antagonist, tropisetron, onto trigeminal subnucleus caudalis/upper cervical spinal cord junction region significantly reduced both formalin-evoked Deep-wide dynamic range unit and pinch-evoked Skin-wide dynamic range unit discharges in non-Complete Freund's Adjuvant and Complete Freund's Adjuvant day 7 groups significantly. The inhibitory effects of tropisetron on pinch-evoked Skin-wide dynamic range unit discharges were prolonged in Complete Freund's Adjuvant day 7 group compared with those in non-Complete Freund's Adjuvant group. The role of central serotonin 3 receptors in trigeminal subnucleus caudalis/upper cervical spinal cord junction region was also tested by orofacial formalin test in Complete Freund's Adjuvant day 7 group. Intracisternal administration of tropisetron decreased the orofacial nocifensive behavior in the late phase evoked by the injection of formalin into the masseter muscle. These results suggest that central serotonin 3 receptors in trigeminal subnucleus caudalis/upper cervical spinal cord junction region are involved in mediating pronociceptive effects in both superficial and deep craniofacial tissues nociception during persistent temporomandibular joint inflammation.     

Induction of autoimmune valvulitis in Lewis rats following immunization with peptides from the conserved region of group A streptococcal M protein

Rheumatic heart disease (RHD) is considered to be an autoimmune disorder mediated by group A streptococcal (GAS) M protein-specific T cells and antibodies that cross-react with cardiac antigens and epitopes of the GAS M protein. In this study, Lewis rats were immunized with a pool of overlapping peptides spanning the conserved region of the GAS M protein in Complete Freund's Adjuvant, followed by immunization with Bordetella pertussis. Controls received adjuvants alone. Spleen-derived lymphocytes from rats immunized with the conserved region peptides proliferated in response to the immunogen and to cardiac myosin. Moreover, histological examination of cardiac tissue from rats immunized with conserved region peptides revealed the presence of inflammatory lesions in both the myocardium and valve tissue indicating a role for GAS M protein-specific autoreactive T cells in the development of cardiac lesions. This study may support the use of the rat model of autoimmune valvulitis to investigate the immunopathogenesis of RHD and possible preventive strategies.     

Experimental autoimmune uveoretinitis (EAU) induced by retinal interphotoreceptor retinoid-binding protein (IRBP): differences between EAU induced by IRBP and by S-antigen

Rats immunized with the retinal interphotoreceptor retinoid-binding protein (IRBP) develop an inflammatory eye disease, "experimental autoimmune uveoretinitis" (EAU). The ocular changes which characterize the EAU induced by IRBP resemble those seen in rats which develop EAU by immunization with another retinal protein, S-antigen (S-Ag). Yet, the two antigens do not cross-react antigenically and the two diseases differ by several features: At low doses (less than or equal to 4 micrograms/rat), IRBP was more uveitogenic in Lewis rats than was S-Ag, inducing disease more reproducibly and with earlier onset time. On the other hand, at higher doses (greater than or equal to 20 micrograms/rat) the disease induced by S-Ag was more severe than that induced by the same doses of IRBP. Rats of various inbred strains differed in their susceptibility to EAU induced by these two antigens. In particular, BN rats were more susceptible to IRBP-induced EAU than to the S-Ag-induced disease, while WF and RCS-rdy+ rats developed severe EAU when immunized with S-Ag but showed minimal or no ocular change when immunized with IRBP.     

In Vitro Effect of Inosiplex on T Lymphocytes. I Influence on T Cells with Receptors for IgG (T γ)

Inosiplex, a complex of inosine and 2-hydroxypropyldimethyl ammonium-4-(acetylamino) benzoate, 1:3 molar ratio, originally developed for antiviral use, is now under wider investigation because of its immunopotentiating properties.

This compound can have some actions on T cells at various stages of differentiation, thus promoting an enhancement of their blastogenic responses to varied mitogenic agents (PHA, Con A, PWM, MLC, tetanus toxoid, and viral antigens).

Our studies demonstrate that under the influence of inosiplex human peripheral blood T lymphocytes bearing Fc IgG receptors have an augmented receptor avidity for SRBC which result in an increased E active rosette formation, and that T cells preincubated with the drug at the appropriate concentrations express more Fc IgG receptors.

Even though Tγ cells exert “in vitro” immunoregulatory properties, the increase in percentage of Tγ lymphocytes do not correlate with a potentiation of the Con A-induced suppressor activity of T cells.

Moreover, the lymphocytes treated with the substance in the absence of Con A exert helper functions, increasing the mitogenic responses of the second culture PHA - treated lymphocytes.

These data appear to suggest a pro-proliferative inosiplex-induced effect which could mask a concomitant suppressor cell induction.     

Crossreactivity with sporozoites, exoerythrocytic forms and blood schizonts of Plasmodium berghei in indirect fluorescent antibody tests with sera of rats immunized with sporozoites or infected blood.

IFA studies are reported using plasmodial antigens from three different stages of the life cycle of Plasmodium berghei: sporozoites (SP); exoerythrocytic schizonts in rat liver (EEF); and parasitized rat erythrocytes (SCH = schizonts). Two series of specific sera were applied: sera from adult rats with a blood-induced infection (series A) and sera from rats immunized against sporozoites by mosquito bites and protected against parasitaemia by chloroquine (series B). In series A antibody titres with all three antigens were seen, but those with SCH were generally the highest. Superinfection with parasitized rat blood did not change the titre. In series B sera, collected from rats after a single exposure to infected mosquitoes, showed only titres with SP from day 3 onwards, but after a second exposure titres to all three antigens developed. Crossreactivity with the heterologous antigens in series B was clearly less than in series A. Anti-P. berghei sporozoite antibodies did not crossreact with P. vivax sporozoites. Rats of series A were resistant to a challenge of parasitized blood and could also inhibit the development of sporozoites. Rats of series B were protected against a challenge of sporozoites but not of infected blood. The results are discussed.     

Monocyte Locomotion in Anergic Chronic Brucellosis Patients: The In Vivo Effect of Ascorbic Acid

In 14 patients suffering from relapsing chronic brucellosis who were anergic to brucella antigens, we have studied peripheral blood monocyte random migration and chemotaxis against non-specific and specific leukoattractants, as well as plasma and monocyte ascorbic acid levels.

We found that all parameters studied, were significantly beneath normal, when compared to normal subjects.

After the oral administration of ascorbic acid at a daily dose of 1gr for 15 conseguetive days, random and directed migration against a non-specific stimulus (casein)returned to normal. Directed migration against disease associated leukoattractants (brucella melitensis and brucella abortus) antigens improved significantly, without reaching normal values.

We concluded that ascorbic acid supplementation might partially restore peripheral, monocyte function and help the monocyte-macrophage system to mount an effective immune response against chronicity of brucella infection.     

Papillary Serous Carcinoma with Basophilic Cytoplasmic Granules

Case History

A 69-year-old white female presented with vaginal bleeding and abdominal distention. A uterine cervical biopsy revealed adenocarcinoma. A laparotomy was performed, with a hysterectomy, bilaterial salpingo-oophorectomy, and multiple omental and abdominal lymph node biopsies. The tumor was present in the endometrium and endocervix with deep myometrial invasion, in both ovaries, numerous peritoneal nodules, and multiple retroperitoneal lymph nodes. Despite postoperative chemotherapy, the patient developed bowel obstruction and died 4 months after surgery. No autopsy was performed

Study the figures without reading the legends and make your diagnosis     

Immunological Properties of a Trypanosomal Lipopolysaccharide and its Effects on the Infection with Trypanosoma (Schizotrypanum) cruzi

A lipopolysaccharide, TCLP, was obtained from culture form of T.(S) cruzi by extraction with phenol-water at 68° C. Unlike the lipopolysac-charides obtained from Gramnegative bacteria, TCLP lacked adjuvant action on antibody formation, did not react with complement and was incapable of sensitizing rabbits for or eliciting the Shwartzman phenomenon, Mice given TCLP showed an initial short period of increased reticuloen-dothelial clearance of colloidal carbon, followed by a 2-day period of significantly depressed clearance.

TCLP had an adverse effect on mice infected with virulent forms of T. (S.) cruzi. Peak parasitemias were significantly increased over those of control animals (given only the parasite). Early mortality also ensued. These effects were found to be independent of the sequence of administration of TCLP and the parasite.

Prior immunization with TCLP did not prevent either the disease or death in mice challenged with virulent T.(S.) cruzi     

Autoantibodies to myeloperoxidase aggravate mild anti-glomerular-basement-membrane-mediated glomerular injury in the rat.

Autoantibodies to myeloperoxidase (MPO) are present in sera from patients with various forms of vasculitis-associated glomerulonephritis. Evidence for a pathogenic role of anti-MPO antibodies has been provided mainly by in vitro studies. We studied the pathogenic role of autoantibodies to MPO in a rat model of mild immune-mediated glomerular injury. Brown Norway rats were immunized with human MPO in complete Freund's adjuvant or with complete Freund's adjuvant alone. At 2 weeks after immunization, rats had developed antibodies to human and rat MPO as detected by indirect immunofluorescence, enzyme-linked immunosorbent assay, and immunoprecipitation. At this time point, rats were intravenously injected with a subnephritogenic dose of 150 micrograms of rabbit anti-rat GBM. Rats were sacrificed at 4 hours, 24 hours, 4 days, and 10 days after antibody administration. Control immunized rats developed mild glomerulonephritis characterized by slight proteinuria at day 10 (14.8 +/- 8.1 mg/24 hours) and moderate intraglomerular accumulation of ED1+ macrophages. Crescent formation, tuft necrosis, and tubular atrophy were not observed in those rats. In contrast, rats immunized with MPO developed severe glomerulonephritis characterized by the early occurrence of severe hematuria, marked proteinuria at day 10 (76.2 +/- 18.2 mg/24 hours), and massive glomerular deposition of fibrin. Complement and rat IgG were present in insudative lesions, but no linear pattern along the glomerular capillary wall was observed. By light microscopy, severe glomerular lesions were found at day 10 consisting of crescent formation and fibrinoid necrosis of capillary loops. In the interstitium, tubular necrosis and atrophy and marked interstitial mononuclear infiltration were found in conclusion, autoantibodies to MPO severely aggravate subclinical anti-GBM disease demonstrating their in vivo pathogenic potential.