bluuebunny/crossref_metadata_2025 huggingface dataset:
Soluble interleukin-2 receptor and soluble CD8 in liver cirrhosis and obstructive jaundice
Authors: WAGNER F , ASSEMI C , LERSCH C , HART R , CLASSEN M
Published: July 2010

Activated lymphocytes secrete soluble interleukin-2 receptor (sIL-2R); CD8-positive lymphocytes secrete soluble CD8 (sCD8). Liver dysfunction in cirrhosis and obstructive jaundice is known to result in depressed cellular immunity. To evaluate whether this is due to real inactivation of the immune system, we measured sIL-2R and sCD8 in the serum of 46 patients with liver cirrhosis, 25 patients with obstructive jaundice, 32 patients with alcoholic liver disease without evidence of cirrhosis, 23 healthy persons and 43 patients with unrelated disease. sIL-2R in patients with cirrhosis (mean ± s.e.m. 1499.140 U/ml) and obstructive jaundice (1517 ± 204) was significantly increased compared with healthy subjects (363 ± 29) and patients with unrelated diseases (685 ± 92); sCD8 was significantly increased in patients with cirrhosis (737 ± 63) but not in patients with obstructive jaundice (419 ± 32) compared with healthy subjects (322 ± 23) and patients with unrelated diseases (375 ± 22). No difference was found between patients with cirrhosis due to alcohol abuse (n= 15) and chronic hepatitis B (n = 6). The Child-Pugh score had no significant influence on the sIL-2R or sCD8 value. In obstructive jaundice, sIL-2R correlated with alkaline phosphatase as marker of cholestasis (r= 0.43). These data show that in spite of the apparent depressed cellular immune defense both in liver cirrhosis and obstructive jaundice there is a general activation of the immune system but the CD8+ cell compartment is only activated in liver cirrhosis. The great changes of sIL-2R and sCD8 in liver dysfunction are important for the interpretation of studies using these serum proteins as markers for immune activation.
Interpretation of blocking activity in maternal serum depends on the equation used for calculation of mixed lymphocyte culture results
Authors: PARK M I , EDWIN S S , SCOTT J R , BRANCH D WARE
Published: July 2010

Immunosuppressive blocking factors in maternal serum are usually determined by inhibition of mixed lymphocyte cultures (MLC), but reports on the importance of these factors for successful pregnancy are conflicting. Here we measured serum blocking activity in men, non-pregnant nulliparous women, non-pregnant multiparous women, women with normal pregnancies, and in women who had had recurrent spontaneous abortions and were treated with leucocyte immunizations. Three different equations were used for calculation of blocking activity: blocking effect index (BEI); stimulation index (SI); and blocking index (BI). By all three methods of calculation, significantly lower levels of blocking activity were noted for men and women compared with pregnant women and multiparae. In the patients with a history of recurrent spontaneous abortions blocking activity as determined by BEI and BI increased into the positive range after treatment with infusions of third-party donor leucocytes in a statistically significant number of women (P < 0.05). However, blocking activity as determined by BEI had a higher predictive value for successful pregnancy than did that determined by BI or SI. Our data suggest that the equation used for calculating BEI is superior to other methods for the determination of blocking activity when monitoring the response to leucocyte immunization in women with recurrent spontaneous abortion. However, these results also cast doubt on the importance of blocking antibodies in histories of recurrent abortion, since pregnancies occurred in the absence and spontaneous abortions occurred in the presence of blocking activity.
Antibody repertoire against culture filtrate antigens in wild house mice infected withMycobacterium bovisBCG
Authors: HUYGEN K , PALFLIET K , JURION F , LENOIR C , VAN VOOREN J-P
Published: July 2010

Wild house mice (Mus domesticus) captured in a Flemish pigsty were infected intravenously with 4 × l06 viable units of Mycobacterium bovis BCG and examined by Western blot analysis for IgG secretion against BCG culture filtrate (CF) antigens. Wild mice showed a marked individual variation in antibody pattern when tested 4, 6 and 8 weeks after infection. Some animals reacted to a wide range of antigens and others only to a limited number. Most wild mice recognized preferentially antigens with molecular weight of 24 kD, 32 kD, 37–38–40 kD, 65 kD and 82 kD, i.e. the major CF antigens known to be recognized by sera from BCG-infectcd inbred laboratory strains, BALB/c, DBA/2, CBA/Ca and C57BL/6. The 32-kD fibronectin-binding protein and the 65-kD heat-shock protein appeared as very immunodominant in wild mice. Furthermore, about 20–25% of the mice reacted strongly with a unique antigen of 35 kD estimated molecular weight, to which the tested inbred laboratory mice did not respond. Monitoring the size of the bacterial population in the spleen indicated that the BCG inoculum did not replicate in wild mice, suggesting that the Bcgr allcle is expressed in this population.
Cellular immunity in experimental Echinococcus multilocularis infection. II. Sequential and comparative phenotypic study of the periparasitic mononuclear cells in resistant and sensitive mice
Authors: BRESSON-HADNI S , LIANCE M , MEYER J P , HOUIN R , BRESSON J L , VUITTON D A
Published: July 2010

Cellular immune responses have been shown to be associated with differential evolutions of E. multilocularis infection in intermediate hosts. A relationship between course of delayed-type hypersensitivity (DTH) against parasitic antigens and receptivity of murine strains has been demonstrated recently. The aim of this study was to correlate resistance and sensitivity to E. multilocularis infection with the phenotypic patterns of cells within the periparasitic granuloma. Evolution of the ratios, macrophages/T lymphocyte and Lyl/Ly2 T lymphocytes, was associated with the receptivity of the strains. Persistence of numerous L3T44+T lymphocytes and low numbers of macrophages and Ly2+T lymphocytes were observed in the ‘resistant’ C57BL.10 mice. Comparison of the results with course of the DTH against E. multilocularis antigens showed that the particular phenotypic pattern observed in resistant mice was associated with a particular profile of DTH after infection. These results and similar observations in human alveolar echinococcosis suggest that cell composition of the periparasitic granuloma might be of crucial importance in controlling the spontaneous development of E. multilocularis larvae in the intermediate host.
Intra-articular immunization induces strong systemic immune response in humans
Authors: TROLLMO C , CARLSTEN H , TARKOWSKI A
Published: July 2010

There is no information available about immunological interactions between the synovial tissue compartment and systemic immunity in health and in disease. The aim of the present study was to evaluate effects of intra-articular immunization on the systemic immune responses in humans. Control subjects were immunized with the same dose of immunogen subcutaneously. Peripheral blood lymphocytes were analysed by spot-ELISA with respect to numbers of immunoglobulin-producing cells and antigen-specific antibody-secreting cells before and 1 week after immunization. Serum and salivary antibody levels were measured by an ELISA before and 14 days after the antigenic exposure. In addition, serum levels of interleukin-6 (IL-6) were analysed before and after immunization. The results indicate that the influenza virus antigen deposited in the joint space induces strong systemic antibody response of IgG, IgA and IgM classes. This response is significantly higher (P < 0.05) than that of control subjects immunized subcutaneously. In contrast, no significant differences were detected between intra-articularly and subcutaneously immunized subjects with respect to mucosal immune responses. Increased serum levels of IL-6 were observed 1–2 weeks after the vaccination in both experimental groups. We conclude that human joints possess very efficient antigen-presenting properties enhancing systemic B cell reactivity.
Soluble HLA class I antigen secretion by normal lymphocytes: relationship with cell activation and effect of interferon-gamma
Authors: BRIEVA J A , VILLAR L M , LEORO G , ALVAREZ-CERMEÑO J C , ROLDÁN E , GONZALEZ-PORQUÉ P
Published: July 2010

HLA class I antigens are thought to be integral membrane proteins. However, soluble forms of these molecules have been detected. Our laboratory has recently shown that the predominant form of these soluble proteins present in human serum, spleen tissue and culture supernatant of activated lymphocytes exhibits molecular weight and structure similar to classical HLA class I antigens, but lacks HLA A or B polymorphic determinants. In the present study, the secretion of such soluble proteins by lymphocytes has been further explored. Phytohaemagglutinin-stimulated normal lymphocytes secrete considerable quantities of soluble HLA (sHLA) class I proteins. This secretion seems to be a general property of lymphocytes, since activation of T as well as B cells by appropriate mitogens equally induce sHLA I secretion. Lymphocytes require RNA and protein synthesis, but not DNA synthesis, for the secretion to occur. Kinetic studies reveal that maximal sHLA I secretion precedes the peak of DNA synthesis by 24 h. In vitro stimulation with antigens or alloantigens also provokes sHLA I secretion. Moreover, this phenomenon has also been detected for in n'ro-aclivated lymphocytes, as enhanced spontaneous sHLA I secretion was observed in cultures of Low-density blastic B and T cells, and of blood lymphocytes obtained from normal subjects who had received a booster immunization 5 days earlier. Interferon-gamma (IFN-γ) increases the expression of membrane-bound class I antigens but does not induce any sHLA I secretion, suggesting that both molecules are under different regulatory mechanisms. Our results indicate that human lymphocytes, upon stimulation, actively secrete considerable amounts of a soluble form of these biologically relevant proteins.
Transforming growth factor-β1 inhibits activation of macrophage cell line RAW 264.7 for cell killing
Authors: HAAK-FRENDSCHO M , WYNN T A , CZUPRYNSKI C J , PAULNOCK D
Published: July 2010

Transforming growth factor β-1 (TGF-β) is a multi-potent immunoregulatory peptide that has effects on numerous cell types. Here we report that human TGF-β inhibits the activation of the macrophagc cell line RAW 264.7 for killing of the L1210 tumour cell line. RAW 264.7 cells, like normal macrophages, require sequential interaction with priming and triggering stimuli for full activation of cytolytic activity. TGF-β inhibits this cytotoxicity in a dose-dependent manner at both the priming and the triggering stage. Addition of as little as 1 ng/ml TGF-β when added with either the priming signal, recombinant interferon-gamma (IFN-γ), or the triggering signal, bacterial lipopolysaccharide (LPS), completely abrogated tumouricidal activity. Incubation with TGF-β also inhibited the morphological changes normally observed in activated RAW 264.7 cells. However. TGF-β was unable to inhibit the cytotoxic activity of RAW 264.7 cells against the target cell line WEHI 164, which is sensitive to tumour necrosis factor. In contrast to the effects on cytotoxic activity, the cytostatic activity of activated RAW 264.7 cells was not inhibited by TGF-β at doses of up to 5 ng/ml. In addition, pretreatment of the LI210 target cells with TGF-β made them refractory to both the cytostatic and cytotoxic effects of RAW 264.7 cells. These data suggest that TGF-β may be an important mediator in the regulation of macrophage tumouricidal activity.
Dose-dependent increase in plasma interleukin-6 after recombinant tumour necrosis factor infusion in humans
Authors: SHERON N , LAU J N , HOFMANN J , WILLIAMS R , ALEXANDER G J M
Published: July 2010

Several studies have shown that the cytokine interleukin-6 (IL-6) is produced in response to tumour necrosis factor (TNF) in vitro. This study examines the in vivo relation between these two cytokines with assays of plasma IL-6 and TNF levels in subjects with chronic hepatitis B undergoing immunomoduiatory therapy with recombinant TNF(rTNF). Plasma IL-6 was detected from 20 min after rTNF infusion with levels peaking after 2–3 h and levels correlated with the dose of rTNF administered (r= 0.67,P=0.004). Peak levels of IL-6 (mean 295, range 266–297 ng/l) were lower than those seen in certain disease states despite the very high peak levels of rTNF (mean 11 750, range 5623-18 620 ng/l). These findings suggest that the very high levels of IL-6 found in certain disease states are not purely the result of circulating TNF. Other factors such as endotoxin or other cytokines may also play a role in determining levels of plasma IL-6.
Isotype-selective abrogation of established IgE responses
Authors: HayGlass K T , STEFURA W
Published: July 2010

Chemically modified allergens have been extensively studied in an attempt to develop materials of increased efficacy and improved safety for use in the immunotherapy of allergic disease. Most of the strategics that have been developed yield products that strongly inhibit de novo IgE responses but have only marginal impact on ongoing IgE responses. We report the virtual abrogation of pre-established murine anti-ovalbumin IgE responses using a glularaldehyde-polymerized ovalbumin preparation (OA-POL) of Mr 3.5 × 107. Secondary IgE responses are inhibited by 97–99% over a period of at least 8 months following three i.p. courses of OA-POL treatment. Administration of five additional ovalbumin [Al(OH)3] booster immunizations over tbis period fails to alter this unresponsive state. The inhibition of antigen-specific IgE responses is isotype specific.
An experimental influenza subunit vaccine (iscom): induction of protective immunity to challenge infection in mice after intranasal or subcutaneous administration
Authors: LÖVGREN K , KÅBERG H , MOREIN B
Published: July 2010

An experimental influenza virus (A/PR/8/34(H1N1)) vaccine was tested and evaluated in mice. The mice were inoculated once or twice intranasally or subcutancously with 1 or 10 μg of iscoins prior to challenge with high doses of live virus. It was demonstrated that two intranasal administrations were as efficient as two s.c. administrations, both routes inducing high levels of antibody and protection against challenge infection. With a one-dose regimen, the s.c. route induced a somewhat higher antibody response than the intranasal route; this might be explained by technical difficulties connected with an intranasal administration.
Tumour necrosis factor production in fulminant hepatic failure: relation to aetiology and superimposed microbial infection
Authors: DE LA MATA M , MEAGER A , ROLANDO N , DANIELS H M , NOURI-ARIA K T , GOKA A K J , EDDLESTON A L W F , ALEXANDER G J M , WILLIAMS R
Published: July 2010

Tumour necrosis factor-alpha (TNF-α), a cytokine derived from macrophages, is considered to bean important endogenous mediator of endotoxic shock. Patients with fulminant hepatic failure are particularly susceptible to infection and the development of multi-organ failure and similarities to endotoxic shock suggest a possible pathogenetic role for TNF in fulminant hepatic failure. In vitro TNF production was therefore investigated serially in 21 consecutive patients with fulminant hepatic failure and in 21 healthy controls. Spontaneous and lipopolysaccharide-stimulated TNF production were elevated in viral-induced fulminant hepatic failure, compared with healthy controls (P < 0.05 and P <0.01, respectively). By contrast, patients with paracetamol-induced fulminant hepatic failure had normal spontaneous and lipopolysaccharide-stimulated TNF production, while those who died had significantly reduced spontaneous TNF production compared with survivors (P < 0.02); this difference was present throughout admission. In this group elevations in TNF production above baseline were associated with Gram-positive bacterial or fungal infection but not Gram-negative bacterial infection. There was no correlation between any of the clinical complications of fulminant hepatic failure and TNF production. These studies indicate that TNF is produced in response to microbial stimuli in fulminant hepatic failure, but do not support a direct role for TNF in the evolution of the clinical complications of fulminant hepatic failure.
Helicobacter pylori induces lymphocyte activation in peripheral blood cultures
Authors: KARTTUNEN R , ANDERSSON G , POIKONEN K , KOSUNEN T U , KARTTUNEN T , JUUTINEN K , NIEMELÄ S
Published: July 2010

Helicobacter pylori-induced, in vitro stimulation of mononuclear cells was characterized by measuring DNA synthesis response, interferon-gamma (IFN-γ) secretion and the number of immunoglobulin-secreting cells. The strength of these responses was measured in 51 subjects comprising 36 dyspeptic patients from the Gastroenterological Unit and 15 members of the laboratory staff. Nineteen subjects had antibodies to H. pylori and 32 did not. The responses were compared with respect to H. pylori antibody status. Positive stimulation of DNA synthesis (stimulation index > 2) was obtained in 96% of the subjects, and the bacterium induced IFN-γ secretion in all of them. The induction of immunoglobulin-secreting cells revealed B cell in vitro stimulation. The antibody-positive subjects had a tendency to synthesize less DNA (P not significant) and secrete less IFN-γ (P < 0.05) in response to H. pylori than did the antibody-negative ones, but the differences were not marked. The results show that the whole H. pylori bacterium stimulates mononuclear cells. The nature of the stimulation is not yet characterized (non-specific versus specific).
Differential upregulation of intercellular adhesion molecule-1 in coeliac disease
Authors: STURGESS R P , MACARTNEY J C , MAKGOBA M W , HUNG C-H , HASKARD D O , CICLITIRA P J
Published: July 2010

The expression of intercellular adhesion molecule-l (ICAM-1) was studied on peroral jejunal biopsies from patients with coeliac disease. The biopsies from untreated patients exhibited greater staining of the superficial lamina propria cells compared with treated patients and controls. A gluten challenge in treated patients produced an altered staining pattern within 2 h. The results demonstrate the role of ICAM-1 expression in coeliac disease, providing further evidence for the role of lamina propria cells in the pathogenesis of this condition.
Lipopolysaccharide from Gram-negative bacteria enhances polyclonal B cell activation and exacerbates nephritis in MRL/lprmice
Authors: CAVALLO T , GRANHOLM N A
Published: July 2010

Depletion of B cells in mice bearing the lymphoproliferation (Ipr) gene reduces lymphoprolifcralion and polyelonal B cell activation (PBA) and attenuates mononuclear cell vasculitis. We sought to verify whether the obverse was true. i.e. whether enhancement of B cell activity might exacerbate the nephritis of MRL /lpr( MRL) mice, a lupus-prone strain. The experimental approach was designed to address three questions: whether naturally occurring PBA in MRL mice could be further enhanced; whether enhanced PBA would exacerbate nephritis: and whether the mechanism of nephritis exacerbation involved interference with mononuclear phagocyte system (MPS) function. To enhance B cell activity, we injected MRL mice with lipopolysaccharide (LPS) from Gram-negative bacteria, a potent B cell activator. To determine whether nephritis was exacerbated, we performed immunopathologic studies and tests of renal function. To verify whether nephritis exacerbation involved impairment of MPS function, we probed the kinetics of immune complex removal from the circulation, their uptake by the liver and spleen, and their localization in kidney tissue. The results indicate that in MRL mice: (i) spontaneous PBA can be enhanced by LPS; (ii) enhancement of PBA by LPS exacerbates nephritis; and (iii) the MPS is already saturated, presumably due to excessive production of endogenous immune complexes. Thus, further increase in immune complex formation due to enhanced PBA by LPS results in increased localization of immune complexes in kidneys and exacerbated nephritis.
Early antibody responses in human schistosomiasis
Authors: EVENGÅRD B , HAMMARSTRÖM L , SMITH C I E , LINDER E
Published: July 2010

Early diagnosis is important when handling patients with acute schistosomiasis. This state is usually more severe in travellers and tourists than in the immune, resident patients. With increased travelling to areas endemic for schistosomiasis, a tool is needed to solve the problem of differential diagnosis due to the non-specific symptoms of the early stages of the disease. Early appearance of antibodies against excretory/secretory antigens of the intestinal tract in the adult worm was seen in six individuals recently infected with Schistosoma mansoni, using an indirect immunofluorescence technique. The antibodies were of IgM, IgG and IgA classes, and of the IgG1, IgG3 and IgA1 subclasses as detected by ELISA using an antigen preparation of adult worm. On immunoblots, using a freeze-dried adult worm antigen, IgG1 and IgG3 antibodies recognized antigens of 32–35 kD. Antibodies against these antigens could thus be a marker of early infection in previously non exposed visitors to endemic areas.
Cells and mediators which participate in immunoglobulin synthesis by human mononuclear cells. II. The mechanism of null cell participation in immunoglobulin synthesis and secretion by B cells
Authors: RICHTER M , TAYLOR S , MACPHAIL S , JODOUIN C A
Published: July 2010

Immunoglobulins were synthesized and secreted by human B cells cultured with T cells with receptors for FcM (TM) helper cells, monocytes, null cells and PWM for 7 days. Immunoglobulin synthesis did not take place if the null cells were omitted from the cultures irrespective of the duration of the culture period. Null cells incorporated into the cultures at only 25% of their optimal concentration did not affect immunoglobulin synthesis markedly by the cultured B cells. However, the number of B cells in the culture could not be diluted without an accompanying marked reduction in immunoglobulin synthesis. The B cells synthesized and secreted significant quantities of immunoglobulin even when the null cells were added as late as day 6 of the 7-day culture whereas no or very little immunoglobulin was synthesized if the B cells were not present from the beginning of the 7-day culture. It was demonstrated that cultured null cells do not transform into B cells and do not attain their immunoglobulin-synthesizing function. Furthermore, cultured B cells do not transform into null cells and do not attain their helper function. The null cells can also be distinguished from the B cells on the basis of cell-surface markers, receptors, and blastogenic responsiveness to phytomitogens. It is concluded that (i) the human circulating B cells require the null cells, in addition to the TM cells, monocytes and PWM, in culture in order to synthesize and secrete immunoglobulin; (ii) the null cell signal that stimulates immunoglobulin synthesis and secretion by the B cells is probably the last signal following the TM helper cell, monocyte and PWM signals received by the B cells; and (iii) the null cells and the B cells constitute distinct lineages of cells.
Molecular size heterogeneity of immunoglobulins in health and disease
Authors: ROBERTS-THOMSON P J , SHEPHERD K
Published: July 2010

The molecular size of serum IgG, IgA and IgM in patients with a variety of monoclonal and polyclonal immune disorders has been determined by a sensitive immunoblotting technique. IgM, IgA and IgG3 paraproteins from patients with B cell lymphoproliferative disorders frequently polymerize with IgA paraproteins demonstrating two polymeric series, the basic unit of the more dominant series being monomeric IgA, and the second consists of a basic unit having a molecular mass of approximately 70 kD heavier than monomeric IgA. The molecular nature of this heavier IgA moiety was shown to be IgA covalently bound with a single moleculeof serum albumin or alpha anti-trypsin and this moiety was also observed in small quantities in sera from healthy subjects. IgM paraproteins, particularly from patients with malignant Iymphoproliferativc disorders, consisted of varying proportions of decamers, pentamers and monomers together with other low molecular weight IgM oligomers. Paraproteins from patients with benign conditions showed less tendency to exist in multiple molecular weight forms. Serum immunoglobulins from patients with polyclonal immune disorder also showed molecular sized heterogeneity. Significantly increased levels of dimeric IgA were observed in patients with alcoholic cirrhosis and Felty's syndrome, while low molecular weight IgM was commonly seen in patients with autoimmune disorders. In these latter disorders monomeric IgM correlated significantly with the serum IgM level suggesting a disorder of assembly of the IgM subunits during an ongoing IgM immune response. We have demonstrated considerable molecular size heterogeneity of serum immunoglobulins both in health and disease and have indicated some possible clinical associations.
Naturally occurring human IgG antibodies to intracellular and cytoskeletal components of human platelets
Authors: PFUELLER S L , LOGAN D , TRAN T T , BILSTON R A
Published: July 2010

Immunoblotting of platelets that have been subjected to SDS-PAGE has revealed that sera from normal individuals contain IgG which binds to many platelet components. This binding was seen with autologous and heterologous platelets using serum of males and of nulliparous females who had not received blood transfusions. Although binding patterns of different sera were not identical, almost all sera caused IgG binding to platelet components of 87–90 kD, 140 kD (identified as vinculin) and 220–240 kD (tentatively identified as talin and actin-binding protein). Purified IgG showed the same binding pattern as whole serum and F(ab')2 fragments retained their ability to bind to many components. The titre of IgG binding in serum was 1:50–1600 while that of alloantibodies to the PIAI antigen was 1:3200. IgG binding components were not secreted when platelets were stimulated and were rarely associated with isolated membranes, but were located either in platelet cytoplasm or cytoskeletons, IgG binding was decreased by absorbing sera with lysed platelets or isolated cytoskeletons, but only slightly with intact platelets. Microaffinity purification of IgG which formed a major band on immunoblots showed that it was antibody with specificity for vinculin or its degradation products. These findings suggest that normal sera contain naturally occurring IgG antibodies with specificity for intracellular platelet antigens and that in some cases their titre approaches that of antibodies of pathological significance.
Depressed lymphocyte transformation and the role of prostaglandins in atopic dermatitis
Authors: JAKOB T , HUSPITH B N , LATCHMAN Y E , RYCROFT R , BROSTOFF J
Published: July 2010

We have shown that peripheral blood mononuclear cells (PBMC) from patients with atopic dermatitis have a reduced in vitro proliferative responsiveness to concanavalin A when compared with non-atopic controls, Addition of the cyclo-oxygenase inhibitor indomethacin caused a significant enhancement of the mitogen response in the patients, indicating a suppressive effect of cyclooxygenase products. We have further demonstrated increased levels of prostaglandin E2 in the supernatants of the PBMC cultures and mereased levels of IgE immune complexes in the sera of the aotpic dermatitis patients and therefore hypothesize that IgE immune complexes may cause increased monocyte production of prostaglandins which in turn appears to be responsible for a reduced lymphocyte proliferation.
Combination chemo-immunotherapy: kinetics ofin vivoandin vitrogeneration of natural killer cells and lymphokine-activated killer cells in the rat
Authors: STEWART L S , SEWELL A F , THOMSON A W
Published: July 2010

Rats received a single high dose of cyclophosphamide (Cy) (150 mg/kg). followed 48 h later (on day 0) by immunization with a T cell-dependent soluble antigen, ovalbumin in Freund's complete adjuvant (FCA). The effect of this treatment on lymphoid cell subpopulalions in the spleen, natural killer (NK) cell and interieukin-2 (IL-2) induced lymphokine-activated killer (LAK) cell activity was examined. Cy (with and without ovalbumin) caused a large relative increase (by day 14) in splenic OX 8+, OX 19-cells with NK morphology. A marked relative increase in fresh NK cell activity was noted after Cy+ovalbumin, but not consistently after Cy alone. Elevated NK activity was Cy dose- and time-dependent, was evident within 7 days post Cy/ovalbumin and persisted for at least 28 days. Pooled splente mononuclear cells (MNC), obtained 14 days after Cy/ovulbumin, lost all cytolytic activity against YAC-I cells when cultured in the absence of human recombinant IL-2 (rIL-2). In contrast, similarly maintained cells from normal rats displayed NK activity higher than normal‘fresh’levels. Upon culture in medium containing 500 U/ml rIL-2, however,‘augmented’NK. activity was equivalent, on a per-cell basis, in both normal and Cy/ovaibumin-pretreuted groups. LAK activity generated in vitro (i.e. against NK.-resistant target cells) was significantly lower in the latter group, and the overall yield of cells was reduced. By day 21 after Cy/ovaibumin, augmented NK activity was significantly greater lhan controls, on a per-cell and total culture yield basis. Moreover, LAK activity was now similar between groups. It is concluded that the chemotherapy/immunization protocol which we have used can greatly enhance NK activity in vitro and that these cells are responsive to induction of LAK activity by IL-2 in ritro.
Reduced natural cytotoxic cell activity in patients receiving cisplatin-based chemotherapy and in mice treated with cisplatin
Authors: POWELL C B , MUTCH D G , KAO M -S , COLLINS J L
Published: July 2010

In mice natural cytotoxic (NC) cells are known to play a role in the rejection of tumours that are sensitive to NC-mediated lysis. We have recently shown that in vitro human and murine tumour cells become more sensitive to lysis mediated by NC cells in the presence of the anti-cancer drug cisplatin. If NC activity plays a role in tumour surveillance in humans, then the ability of NC cells to eliminate tumours in patients treated with cisplatin would not only be dependent on cisplatin increasing the sensitivity of tumours to NC mediated lysis, but would also depend on the maintenance of high levels of NC activity during chemotherapy. Here we report that patients receiving chemotherapy that included cisplatin showed a time-dependent reduction in NC activity. NC activity was normal I day after treatment; however, 15 days after treatment patients had an eight- to 16-fold reduction in NC activity that returned to normal levels by day 21. The reduction in NC activity of patients was coincident with a reduction in circulating monocytes. Mice treated with only cisplalin showed a similar reduction in NC activity. Mice treated with cisplalin had a reduced level of NC activity that was first apparent 8 days after treatment, reached a nadir on day 15, and returned to normal levels on day 22.
Antibodies to mouse laminin in patients with systemic sclerosis (scleroderma) recognize galactosyl (αl-3)-galactose epitopes
Authors: GABRIELLI A , CANDELA M , RICCIATTI A M , CANIGLIA M L , WIESLANDER J
Published: July 2010

Employing radioimmunoinhibition assays with distinct oligosaccharides as inhibitors, this study demonstrates that the epitope recognized on mouse laminin by sera from patients with systemic sclerosis (scleroderma) is a terminal galactosyl (α1–3)-galactose disaccharide. The reaction with this α-digalactose was further confirmed when the sera were tested in radioimmunoassay (RIA) binding assay and in ELISA with synthetic galactose α1–3 galactosc coupled to human serum albumin. The circulating antibody appeared restricted to the IgG class and mostly to the subclass IgG3 and IgG4. Antibodies with the same specificity can be found in patients with American cutaneous leishmaniasis and Chagas disease: however, whilst in these diseases the antibody production is triggered by antigenic determinants present on the surface of the parasites, the events eliciting their appearance in systemic sclerosis are unknown.
Heart-specific autoantibodies can be eluted from the hearts of Coxsackievirus B3-infected mice
Authors: NEUMANN D A , LANE J R , LAFOND-WALKER A , ALLEN G S , WULFF S M , HERSKOWITZ A , ROSE N R
Published: July 2010

This study was undertaken to determine if immunoglobulin G (IgG) antibodies could be eluted from the hearts of mice with Coxsackievirus B3-induced autoimmune myocarditis and to characterize the immunorcactivity of any elutable autoantibodies. Susceptible (A/J) and resistant (B10.A) mice were administered the virus or the control treatment and killed at various times after treatment. Acid eluates from pooled heart tissue from each treatment group and each time were tested for IgG reactivity with normal heart tissue by immunohistochemistry and with normal heart extracts by Western immunostaining. Eluates from infected A/J mice reacted strongly with syngeneic heart and modestly with syngeneic skeletal muscle tissue. Eluates from infected B10.A or control mice of either strain exhibited little reactivity with either tissue. Tissue reactivity was similar when allogeneic tissue was used as the substrate. Eluates from infected A/J mice recognized the heavy chain of cardiac myosin and several other cardiac antigens by Western immunostaining while eluates from the other treatment groups exhibited little or no reactivity with any normal heart constituents. These results indicate that in vivo IgG deposition occurs in the hearts of mice with post-infectious autoimmune myocarditis and that the specificity of these antibodies is similar to that reported for serum from animals with this disease. The mechanism(s) leading to myocardial IgG deposition and its possible role in pathogenesis remain to be elucidated.
Inhibition of autoimmune diabetes in NOD mice with serum from streptococcal preparation (OK-432)-injected mice
Authors: SEINO H , SATOH J , SHINTANI S , TAKAHASHI K , ZHU X P , MASUDA T , NOBUNAGA T , SAITO M , TERANO Y , TOYOTA T
Published: July 2010

We have recently reported that systemic and chronic administration of recombinant tumour necrosis factor alpha (TNF-α), as well as streptococcal preparation (OK-432), inhibits development of insulin-dependent diabetes mellitus (IDDM) in NOD mice and BB rats, models of IDDM. In this study we examined whether serum containing endogenous TNF induced by OK-432 injection could inhibit IDDM in NOD mice. Treatment twice a week from 4 weeks of age with OK-432-injected mouse serum, which contained endogenous TNF (75U), but not IL-1, IL-2 and interferon-gamma (IFN-γ) activity, reduced the intensity of insulitis and significantly inhibited the cumulative incidence of diabetes by 28 weeks of age in NOD mice, as compared with the incidence in non-treated mice (P<0.01) and in mice treated with control serum (P<002). This inhibitory effect of the serum was diminished, although not significantly, by neutralization of serum TNF activity with anti-mouse TNF antibody. In the mice treated with the serum from OK-432-injected mice, Thy-1.2+ or CD8+ spleen cells decreased (P<0.01) and surface-Ig+ (S-Ig+) cells increased (P<0.05), whereas the proliferative response of spleen cells to concanavalin A (P< 0.01) and lipopolysaccharide (P<0.05) increased. The results indicate that the inhibition by OK-432 treatment of IDDM in NOD mice was partially mediated by serum factors including endogenous TNF.
Antibodies toMycobacterium tuberculosis-specificepitopes in lepromatous leprosy
Authors: BOTHAMLEY G , BECK J S , BRITTON W , ELSAGHIER A , IVANYI J
Published: July 2010

Sera from patients with leprosy or tuberculosis and healthy subjects have been analysed for the presence of antibodies to four species-specific mycobacterial epitopes, four different viruses and five autoantigens. Antibodies to the Mycobacterium leprae-specific 35-kD protein and phenolic glycolipid I epitopes were not present in patients with active pulmonary tuberculosis. In contrast, antibody levels to species-specific epitopes of the 38-kD and 14-kD antigens M. tuberculosis were significantly elevated in patients with lepromatous leprosy. Neither of the two antigens is cross-reactive with M. leprae at the B cell level. However, it was considered that cross-reactive helper T cells could recall the response of M. tuberculosis-specific memory B cells, which had been primed through prior self-healing tuberculous infection. As an alternative explanation, the possible role of polyclonal B cell stimulation was considered. This seemed unlikely, however, since: (i) antibody levels to autoantigens, except anti-smooth muscle, were not elevated, and (ii) antibody levels to four distinct viruses, unlike those to all mycobacterial epitopes, showed no correlation with litres, to M. tuberculosis-specific epitopes.
Suppression of human lymphocyte responses to specific and non-specific stimuli in human onchocerciasis
Authors: ELKHALIFA M Y , GHALIB H W , ALLA T DAFA , WILLIAMS J F
Published: July 2010

Characterization of in vitro lymphocyte responsiveness was performed on selected groups of onchocerciasis patients from Sudan and Sierra Leone. These patients manifested a very broad range of clinical signs and showed widely divergent parasite infection intensities. Lymphocyte proliferative responses to soluble Onchocerca volvulus antigen (sAg) were poor in infected persons; mitogen and PPD responses were maintained in the normal range in one group of patients from southwestern Sudan, but were profoundly depressed in a group from N.E. Sudan. Proliferative responses and interferon-gamma (INF-γ) secretion were very significantly depressed in the presence of live microfilariae of O. volvulus or secretions/excretions (S/E) from microfilariae (mf) or from female, but not male, adult parasites. Lymphocyte responses were maintained near normal when exogenous IL-2 was added to these cultures. The results indicate that O. volvulus infection and its clinical consequences arc not consistently associated with systemic deficits in immune responsiveness. However, suppression of lymphocyte reactivity by mf and S/E in vitro suggests that direct parasite intervention in host cell responses could be taking place in vivo, perhaps at the local micro-environment level, mediated by effects on cytokine production.
T cell receptor γδ bearing cells are decreased in the peripheral blood of patients with atopic diseases
Authors: SCHAUER U , DIPPEL E , GIELER U , BRÄUER J , JUNG T , HEYMANNS J , RIEGER C H L
Published: July 2010

The biological role of T cell receptor (TCR) γδ bearing cells is currently not fully understood. Recently, a monoclonal antibody (TCRδ1) reacting against the whole molecule became available which facilitates the direct analysis of TCR-γδ+ cells. We studied 11 children with atopic dermatitis, 20 children with atopic asthma, 18 adults with atopic dermatitis and 38 healthy age matched controls aged 4–51 years. Lymphocytes were isolated from heparinized peripheral blood and the proportion of TCR-γδ+ lymphocytes was determined by FACS analysis. Patients with atopic diseases yielded a significantly (P<0.01) lower proportion of TCR-γδ+ cells compared with normal controls (median 4.8%versus 7.1%). The percentage of TCR-γδ+ cells showed an age-dependent decline in both the patient group (r=—0.49, P<0.01) and the control group (r=–-0.40, P<0.01). In addition, the proportion of cells which expressed CD8, TCR-γδ or CD4, TCR-γδ simultaneously was determined by double labelling immunofluorescence. Whereas CD4+, TCR-γδ+ cells could be identified in only a few individuals, CD8+, TCR-γδ+ cells were found in nearly all controls (median 2.4%, range 0.0–10.8%); atopic patients displayed significantly (P < 0.01) lower proportions of CD8+, TCR-γδ+ cells.
Vascular deposition of complement-split products in kidney allografts with cell-mediated rejection
Authors: FEUCHT H E , FELBER E , GOKEL M J , HILLEBRAND G , NATTERMANN U , BROCKMEYER C , HELD E , RIETHMÜLLER G , LAND W , ALBERT E
Published: July 2010

Complement activation in 73 renal transplant biopsies was investigated by indirect immunoperoxidase staining using MoAbs reactive with complement-split products. Intense deposition of complement fragments C4d and C3d in peritubular capillaries, indicating activation of the classical pathway, could be detected in the majority of transplanted kidneys with cell-mediated rejections. Abundant deposition of complement-split products was observed in 22 early biopsies from patients with high ‘immunological risk’ (i.e. previous, rejected transplants and/or circulating antibodies against HLA-antigens). Despite negative results in the crossmatch before transplantation and paucity of immunoglobulins in transplant biopsies, antibodies directed against endothelial cell antigens should be considered as a possible cause of classical complement activation.
Peripheral blood neutrophils in inflammatory bowel disease: morphological evidence ofin vivoactivation in active disease
Authors: MCCARTHY D A , RAMPTON D S , LIU Y-C
Published: July 2010

Morphological evidence of activation in vivo of circulating neutrophils in patients with inflammatory bowel disease (IBD) was sought by quantitative light microscope examination of toluidine bluestained leucocyte preparations made from peripheral venous bloods that had been fixed immediately ex vivo. The proportion of spherical (unactivated) circulating neutrophils was reduced in active Crohn's disease (73%; 46–96 (median; range), n= 11) compared with inactive Crohn's (90%; 45–99; n= 18, P<0.01) and normal subjects (94%; 44–98; n=13, P<0.05). There tended to be fewer spherical neutrophils in active ulcerative colitis (77%; 13–96; n= 17) than in quiescent colitis (88%; 57–99, n= 13, P<0.1) or normal subjects (P<0.05). Activated neutrophils occur in the circulating pool of patients with active IBD and can be detected by light microscopy of peripheral venous blood leucocyte preparations.
Cytotoxicity against human peripheral blood mononuclear cells and T cell lines mediated by anti-T cell immunotoxins in the absence of added potentiator
Authors: FISHWILD D M , STASKAWICZ M O , WU H-M , CARROLL S F
Published: July 2010

Several in vitro assays have indicated that anti-T cell immunotoxins (IT), composed of monoclonal antibodies (MoAbs)conjugated to ricin A chain (RTA), are maximally effective against T cells only in the presence of potentiators. It was thought that such IT might not be sufficiently cytotoxic to deplete T cells in vivo upon administration to patients. Therefore, we have re-evaluated the in vitro assays and report herein that even with a short exposure lime (2 h), the two anti-T cell IT, H65-RTA (anti-CD5 MoAb coupled to RTA) and 4MRTA (anti-CD7 MoAb coupled to RTA30), were specifically cytotoxic for peripheral blood mononuclear cells(PBMC)in the absence of potentiators. Moreover, as has been reported for IT when tested against T cell lines, prolonging the exposure lime of the IT with PBMC from 2 h to as long as 90 h, without added potentiators, enhanced their cytotoxicity from 2- to 40-fold. In contrast, most T cell lines were more sensitive to IT in the presence of potentiator, and IT cytotoxicity was much less enhanced by prolonging the exposure time. Thus, T cell lines may not serve as accurate models to determine the efficacy of IT against PBMC in vitro or in vivo. We conclude that IT-induced cytotoxicity of PBMC can be demonstrated in vitro at pharmacologically achievable concentrations in the absence of added potentiators.
Defective control of Epstein-Barr virus-infected B cell growth in patients with X-linked lymphoproliferative disease
Authors: YASUDA N , LAI P K , ROGERS J , PURTLO D T
Published: July 2010

We studied the cellular function and lymphokine production of T cells from patients with X-linked lymphoproliferative disease (XLP) when activated by the challenge with Epstein-Barr virus (EBV) infection. We used an assay system in which T cells were stimulated with membrane antigens of autologous EBV-infected B lymphoblastoid cell lines (B-LCL) and we examined cellular and humoral factors derived from the stimulated T cells which control the growth of EBV-inlected B-LCL. Immunoglobulin secretion from the autologous B-LCL was suppressed with radiosensitive suppressor cells in the patients with XLP. The degree of suppression was correlated with the immunoglobulin levels in the serum of the patients with acquired hypogammaglobulinaemia (P<0.05). In addition, T cells from the patients with XLP failed to produce interferon-gamma (IFN-γ) (P < 0.001). Moreover, the T cell supernatants from the patients with XLP were less potent to inhibit the B-LCL growth. This diminished inhibition of the B-LCL growth was correlated well with the decreased concentration of IFN-γ in the T cell supernatants. These findings suggest that suppressor cells may be activated in the patients with the hypogammaglobulinaemia phenotype of XLP. but the frequent development of B cell lymphoma in hypogammaglobulinaemia indicate that immunoglobulin suppression may not exert enough pressure on the in vivo growth of EBV-infected B cells. The defective secretion of IFN-γ may be, at least partially, responsible for the abnormal cytotoxic T cell and natural killer activities found in the patients with XLP, and may indicate the clinical evaluation about the preventive injection of IFN-γ against the development of malignant lymphoma.
Human cartilage is degraded by rheumatoid arthritis synovial fluid but not by recombinant cytokines in vitro
Authors: HOLLANDER A P , ATKINS R M , EASTWOOD D M , DIEPPE P A , ELSON C J
Published: July 2010

Rheumatoid arthritis (RA) synovial fluid (SF) stimulated significant loss of glycosaminoglycans (GAG) from normal and pathological human cartilage biopsies over 2 days as compared with normal human serum. By contrast, 15 RA SFs failed to degrade killed normal cartilage, and degraded killed RA cartilage less effectively than living RA cartilage. Four RA SFs were treated with neutralizing anti-cytokine antisera prior to incubation with normal cartilage. The degrading effects of two of the fluids were reversed by anti-interleukin-1α (IL-1α) while degradation by the third and fourth fluids were reversed by anti-intcrleukin-1β (IL-1β) and anti-tumour necrosis factor-alpha (TNF-α). respectively. However, recombinant human IL-1α. 1L-1β. TNFα or a combination of all three cytokines had no degrading effect in this 2-day culture system. It is concluded that RA SF degrades cartilage by a mechanism involving a synergistic interaction between cytokines and some other component of SF.
Cytomegalovirus-specific antibodies to an immediate early antigen and a late membrane antigen and their possible role in controlling secondary cytomegalovirus infection
Authors: VAN ZANTEN J , VAN DER GIESSEN M , VAN DER VOORT L H M , VAN SON W J , VAN DER BIJ W , THE T H
Published: July 2010

In 24 renal transplant recipients who had a secondary cytomegalovirus (CMV) infection, the magnitude and development of CMV-specific antibodies directed against two different antigens were studied in relation to the presence of CMV-immediate early antigen-positive peripheral blood leucocytes (CMV antigenaemia). These antibodies were measured in an antigen-capture ELISA using two monoclonal antibodies: one directed against the major immediate early antigen (IEA) and a second one directed against the CMV-encoded glycoprotein B (gB). A statistically significant inverse relationship between the level of anti-IEA antibodies present at the time of transplantation as well as the magnitude of the increase of these antibodies during CMV infection and the maximum number of IEA-positive cells during infection was shown. In contrast, both anti-gB and anti-total CMV antibodies did not give any correlation with the CMV antigenaemia. This may indicate that the anti-IEA immune response plays a role in defence mechanisms against a CMV infection.