Lymphocyte Transformation Activity Research Articles

Systems pharmacology reveals the mechanism of Astragaloside IV in improving immune activity on cyclophosphamide-induced immunosuppressed mice

Ethnopharmacological relevanceMyelosuppression, also known as bone marrow suppression (BMS), is a pathological phenomenon of the decrease in the production of blood cells and further lead to immune homeostasis disorder. Astragalus mongholicus Bunge (AM, checked with The World Flora Online, http://www.worldfloraonline.org, updated on January 30, 2023) is a traditional Chinese medicine with efficacy of tonifying Qi and strengthening body immunity in thousands of years of clinical practice in China. Astragaloside IV (AS-IV) is a major active ingredient of AM, which plays an important role in regulating immune system through different ways. Aim of the studyThis study was aimed to investigate the protective effect and mechanism of AS-IV on macrophages in vitro and cyclophosphamide (CTX)-induced immunosuppressive mice in vivo, and to provide experimental basis for the prevention and treatment of AS-IV in myelosuppression. Materials and methodsBased on network pharmacology and molecular docking technology, the core targets and signaling pathways of saponins of AM against myelosuppression were screened. And then, the immunoregulatory effect of AS-IV on RAW264.7 cells was investigated by cellular immune activity and cellular secretion analysis in vitro. In this way, the effects of AS-IV on the main potential targets of HIF-1α/NF-κB signaling pathway were analyzed by qRT-PCR and Western blot methods. Furthermore, comprehensive analysis of the effects of AS-IV against CTX-induced mice were conducted on the basis of immune organs indices analysis, histopathological analysis, hematological analysis, natural killer cell activity analysis and spleen lymphocyte transformation activity analysis. In order to further verify the relationship between active ingredients and action targets, drug inhibitor experiments were finally conducted. ResultsAS-IV, as a potential anti-myelosuppressive compound, was screened by systematic pharmacological methods to act on target genes including HIF1A and RELA together with the HIF-1α/NF-κB signaling pathway. Further studies by molecular docking technology showed that AS-IV had good binding activity with HIF1A, RELA, TNF, IL6, IL1B and other core targets. Besides, cellular and animal experiments validation results showed that AS-IV could enhance the migration and phagocytosis of RAW264.7 cells, and protect the immune organs such as spleen and thymus together with bone tissues from damage. By this means, immune cell function including spleen natural killer cell and lymphocyte transformation activity were also enhanced. In addition, white blood cells, red blood cells, hemoglobin, platelets and bone marrow cells were also significantly improved in the suppressed bone marrow microenvironment (BMM). In kinetic experiments, the secretion of cytokines such as TNF-α, IL-6 and IL-1β were increased, and IL-10, TGF-β1 were decreased. The key regulatory proteins such as HIF-1α, NF-κB, PHD3 in HIF-1α/NF-κB signaling pathway were also regulated in the results of upregulated expression of HIF-1α, p–NF–κB p65 and PHD3 at the protein or mRNA level. Finally, the inhibition experiment results suggested that AS-IV could significantly improve protein response in immunity and inflammation such as HIF-1α, NF-κB and PHD3. ConclusionAS-IV could significantly relieve CTX-induced immunosuppressive and might improve the immune activity of macrophages by activating HIF-1α/NF-κB signaling pathway, and provide a reliable basis for the clinical application of AS-IV as a potentially valuable regulator of BMM.

Exploring the potential mechanism of Yixiang capsule immune-enhancing function based on Network Pharmacology, Molecular docking, and Animal function experiment.

The Yixiang capsule with traditional Chinese herbs Cistanches Herba, Hedysarum multijugum Maxim,Fructus Ligustri Lucidi,and Fructus lycii are the main raw materials in a modern herbal medicine preparation containing corresponding bioactive ingredients after extraction and processing. This study aimed to investigate the immune-enhancing function of the Yixiang capsule and explore its potential mechanism. After oral administration of the Yixiang capsule to mice for 30 days, the spleen lymphocyte transformation activity, NK cell activity, DTH, macrophage phagocytosis, HC50 value, amounts of antibody-producing cells, carbon particle clearance rate, spleen, and thymus index were determined to evaluate the regulation effect of the Yixiang capsule on immune function. The potential mechanism underlying the immune-enhancing function of the Yixiang capsule was studied based on network pharmacology and molecular docking. The results showed that the Yixiang capsules can obviously increase the HC50 values in all groups, significantly enhance the DTH reaction of mice, the carbon clearance function in medium and high dose groups,and the phagocytic function of macrophages in high dose groups, but had no significant effect on NK cell activity, T lymphocyte proliferation, body weight and immune organ weight of mice. A total of 78 compounds of the Yixiang capsule may affect 123 metabolic pathways and 1022 GO terms by regulating 232 targets, such as the cancer signaling pathway ad the signal pathways of response to lipopolysaccharide, and the key targets predicted could stably bind to the core active components of the Yixiang capsule. These results suggested that the Yixiang capsule could enhance the immune function of mice and have the characteristics of a multi-component and multi-target synergistic effect on the enhanced immunity function, which provides the basis for the follow-up study.

The immune system ofthe ferret (Mustela putorius furo) - Areview.

The basic information dealing with the anatomy ofthe ferret's immune system, cross-reactivity oftheferret leukocytes with polyclonal and monoclonal antibodies invitro and immune response tothe mitogens and various infections are presented. The leukocyte numbers inthe peripheral blood inthe ferrets are lower compared toother species and only one subclass ofIgG has been identified inferrets sofar. Lymphocytes make up 12-67% ofall theleukocytes inthe peripheral blood ofthe healthy adult ferrets. Lymphocyte subpopulations are similar toother mammals and include T-and B-lymphocytes. T-lymphocytes differentiate into helper (Th) lymphocytes and cytotoxic (Tc) lymphocytes. Currently, ferret granulocytes (CD11), B-lymphocytes (CD79α), T-lymphocytes(CD3), Th-lymphocytes (CD3, CD4), Tc-lymphocytes (CD3, CD8), and CD30, CD45 subpopulations are detected withtheuse ofanumber ofpolyclonal aswell aswith monoclonal antibodies. Inalymphocyte transformation assay, themitogen response ofthe peripheral blood mononuclear cells toconcanavalinA(ConA), phytohaemagglutinin (PHA), and pokeweed mitogen (PWM) isthe greatest atday2, 2 and 3, respectively. Serious lymphopenia isobservedinferrets during adistemper infection. Asignificant decrease inthe lymphocyte transformation activity isobserved onday5 and reaches amaximal decrease ondays 8-11, with full recovery ondays 23-30 after theinoculation oflaboratory ferrets with the distemper virus. Ferrets have also been used instudies related tothe function oftheimmune system inHelicobacter pylori infections, Crohn's disease and bronchial asthma.

Study on the Enhancement of Immune Function of Astaxanthin from Haematococcus pluvialis.

This study was aimed at investigating the effect of astaxanthin on the immune function and its safety in mice. It was administered once daily at low, medium and high doses (4.2, 8.35, 16.70 mg/kg BW) to mice for 30 days. Subsequently, the spleen and thymus index, spleen lymphocyte transformation activity, delayed allergy reaction, amounts of antibody-producing cells, half-hemolytic value HC50, carbon particle clearance rate, macrophage phagocytosis, and natural killer cell (NK) activity were determined. Acute oral toxicity and genotoxicity tests were conducted to evaluate the safety of astaxanthin. Compared with the control group, medium and high doses of astaxanthin significantly increased the proliferation and transformation activities of spleen lymphocytes, activities of antibody-producing cells, serum hemolysin levels, and carbon particle clearance rate in mice (phagocytic index). High doses significantly improved delayed allergy reaction and NK cell activity. Results of acute oral toxicity and genotoxicity tests were negative. Gross anatomical observations and histopathological examination showed no abnormal changes associated with the treatments. In the article, it is confirmed that astaxanthin treatments significantly improve immune functions and show no toxic effects in the experimental doses.

Immunological and pathological adverse effects of avian influenza virus subtype H9N2 infection in aflatoxicated-broiler chickens

Aflatoxin B1 (AFB1) is a metabolic product of the Aspergillus spp. of molds, which grow on several feedstuffs stored in hot moist conditions. It is one of the immunosuppressive agents that might influence the pathogenesisof avian influenza virus (AIV) subtype H9N2 in broilers, which can exacerbate the disease outcomes. The immunological, biochemical and pathological adverse health effects of an interaction between low levels of dietary aflatoxins (AFs) and H9N2 infection in broiler chickens were investigated. One hundred and eighty of unvaccinated 1-day-old COBB chicks were, therefore, raised for 35 days in the following treatment groups: control, AFs, AFs+H9N2, and H9N2. AFs in the basal diet was added at 200 ppb starting from the first day of age, while H9N2 virus was intra-nasally installed at a dose of 100 μl of 106 EID50/bird of allantois fluid at 23rd day. Humoral and cell-mediated immune responses were evaluated. Evidence of H9N2-AIV viral shedding was also detected. It has been observed that concurrent exposure of AFs and H9N2 virus negatively affected chicken performance traits i.e. lowered feed intake and body weights with exaggerated respiratory and digestive disturbances, and 20% mortality rate. Ten days’ post H9N2 infection, significant (p≤ 0.05) increment in serum transaminases (AST and ALT) and falling in cell-mediated immunity i.e. total leukocyte count, lymphocyte transformation activity and macrophage phagocytic activity were detected. Additionally, AFs+H9N2 significantly (p≤ 0.05) lowered H9N2-HI titers (5.5 Log2) than H9N2 alone (6.3 Log2). Pathologically, aflatoxicated chickens showed hydropic degeneration, hepatocytic vacuolation and necrosis of liver tissues with nephrosis and urates deposition in ureters, as well as bursal and thymic lesions, which were potent in H9N2–inoculated chickens. AFs exposure increased the incidence and titer of H9N2 viral shedding. It could be concluded that dietary contamination with AFs even at very low levels has explanatory effect in H9N2–inoculated broilers, and vice versa.

Streptococcal Pyrogenic Exotoxin A, Streptolysin O, Exoenzymes, Serotype and Biotype Profiles of Streptococcus pyogenes Isolates from Patients with Toxic Shock Syndrome and other Severe Infections

The determination of protein M and T serotypes, biotypes and pyrogenic (erythrogenic) exotoxin A (SPE A), streptolysin O (SLO), streptokinase (SK), hyaluronidase (HA) and cysteine proteinase release by 212 S. pyogenes isolates from patients with severe invasive group A streptococcal (GAS) infections, among them 74 cases of streptococcal toxic shock syndrome (STSS) has been investigated. M1 or M3 serotypes were expressed by 25% of the isolates (53/212), whereas 59% (125/212) belonged to 15 other different serotypes and 16% (34/212) were untypeable. Of the 74 isolates from STSS patients, 42% (31/74) expressed M1 and to a lesser extent M3 serotypes versus 19% of the non STSS isolates (26/138). Among the ten different biotypes known, biotypes 1 and 3 were prevalent, particularly the former in the case of STSS isolates. SPE A was detectably produced by about 25% (54/212) of the strains. However, as high as 40.5% of the STSS isolates (30/74) versus 17.4% of non STSS isolates (24/138) released SPE A. Moreover, 67% of the SPE A producing strains were of serotype M1 or M3. SK and HA were released by 71% and 10% of the isolates respectively. All strains released SLO (4 to 256 HU/ml) and 85% cysteine proteinase. No relationship between toxin or enzyme titer and the type of disease or clinical origin of the strains was found. Culture supernatants of all isolates showed moderate to high lymphocyte transforming activity with index values ranging from 14.5 to 50.3 including those strains which did not release detectable amounts of SPE A suggesting that SPE C and other mitogenic factor(s) are released by the isolates investigated.

Identification and biological activity of lectins of different subunit composition isolated from Phaseolus vulgaris L var athropurpurea

AbstractProtein concentrate, albumin (AF) and globulin fractions (GF) (60.2, 42.3 and 69.7% of protein content, N × 5.40, respectively) were obtained from raw seeds of Phaseolus vulgaris var athropurpurea (PHVa) cultivated in northern Spain. SDS‐PAGE analysis revealed that the isolectins were present solely in the AF while the GF was entirely lectin‐free. Affinity chromatography and SDS‐PAGE were used to separate and identify lectins of different subunit composition: E4 + E3L, E2L2, El3 and L4 (in g: 0.12, 0.03, 0.03 and 0.44, respectively, from 2.9g of PHVa meal). In this cultivar the amount of E4 was negligible. With the methodology followed in this study the L4 isolectin could not be isolated from the AF and it was determined by densitometry. The biology activities of these PHVa phytohaemagglutinins—erythroagglutinating and lymphocyte transformation activities—were assessed in rat and human cells. There was a direct correlation between the erythroagglutinating activity and the E subunit composition of the isolectins using rat red blood cells. The lymphocyte transformation activity of the isolectins with human peripheral blood lymphocytes was correlated with the L‐subunit content in the lectins. However, the relationship between lymphocyte transformation activity and lectin profile with rat lymphatic node lymphocytes was not as straightforward as expected.

The Characterization of Two New Low Molecular Weight Proteins (LMPs) from Streptococcus pyogenes

Two novel extracellular mitogenic substances were isolated from Streptococcus pyogenes strain NY-5 and characterized. The purification steps involved an initial enrichment of the proteins from culture supernatant by silica gel adsorption, followed by ion exchange chromatography and gel filtration. The purified materials were homogeneous in SDS-PAGE, showed estimated molecular weights of 12 kD and isoelectric points of 4.7 and 4.3, respectively. Both proteins (LMP-12k-4.3pI and LMP-12k-4.7pI) demonstrated lymphocyte transformation activity at a concentration of 0.1 microgram/ml. The LMP-12k-4.7pI showed a 69.2% homology of the amino acid sequence with that of a phosphocarrier protein of Staphylococcus aureus and with a total identity in the active centre. The same protein was also isolated from streptococcal group C strain H46A with an N-terminal amino acid sequence being identical. The LMP-12k-4.7pI demonstrated biochemical properties identical with those of the earlier described streptococcal pyrogenic exotoxin type D. The LMP-12k-4.3pI did not show such a clear relation to other functional proteins.

Epstein‐barr virus‐infected b cells persist in the circulation of acyclovir‐treated virus carriers

In this study, infectious Epstein-Barr virus (EBV) shedding in the oropharynx and numbers of virus-infected B cells in the blood have been monitored in long-term virus carriers receiving acyclovir (ACV) therapy for herpes zoster. Eleven patients on oral ACV were followed prospectively before, during and for 2 weeks after treatment. As expected, the low levels of EBV shedding in these virus carriers (measured as cord-blood lymphocyte transforming activity in throat washings) were eliminated during the period of ACV treatment and returned at later times. Over the same period, however, the frequency of virus-infected B cells in the blood (measured by spontaneous transformation in limiting dilution assay) remained completely unchanged. Regression assays showed that these same patients had normal levels of EBV-specific cytotoxic T-cell immunity, so that the in vivo persistence of virus-infected B cells could not be ascribed to a defect in T-cell surveillance. We infer that the in vivo half-life of the virus-infected B-cell pool in long-term virus carriers is measured in months rather than days. We further suggest that such persistence requires a novel form of virus:B-cell interaction distinct from the type of "latent" infection displayed by in vitro-transformed cells.

Studies on tests of in vitro lymphocyte transformation to streptococcal T antigens in asthmatic children

Activity of lymphocyte transformation (LTF) to streptococcal T antigens was studied in 7 asthmatic children and normal subjects included 21 children and 12 neonates (cord blood).T antigens used were Ti, T4, T6, T8, T12 and T22.The results are summarized as follows.1.In unstimulated lymphocyte cultures, no significant difference was found out between asthmatic (mean, 74.9±28.0 cpm) and normal children (mean, 83.5±43.3 cpm) in 3H-thymidine uptake.2.The LTF activity to these T antigens in asthmatic children was significantly higher than that of normal subjects.The stimulation index (SI), especially, to T12 antigen in asthmatic children (mean SI=132.2) revealed remarkable elevation as compared with that in normal children (mean SI=4.5).

Cellular immune response to infection with respiratory syncytial virus and influence of breast-feeding on the response.

Virus-specific lymphocyte transformation (LTF) activity in vitro was examined in 78 infants with various forms of illness due to respiratory syncytial virus (RSV) infection. In 73 subjects with lower respiratory tract disease, significant LTF activity was often detected within one week after onset of initial symptoms, and responses characteristic for each clinical form were observed in the subsequent rise. Thus, mean activity in subjects with tracheobronchitis increased gradually, with the maximum response being detected at the fourth week after the onset of illness. In subjects with pneumonia the response was rather low during the first week of illness, and then a sharp increase of activity was observed at the second week. Although patients with bronchiolitis elicited similar levels of the activity until the first week, the response was significantly suppressed during the subsequent two weeks. The response at the second week in this group was significantly lower than those of corresponding specimens obtained from patients with pneumonia (p less than 0.01) or tracheobronchitis (p less than 0.05), suggesting a close association of the responsiveness with underlying mechanisms of bronchiolitis induced by RSV. The present study further indicated a possibility that breast-feeding of RSV-infected infants may alter levels of the LTF activity in these subjects.

Cell-mediated immunity to cytomegalovirus in immunocompromised patients: effect of radiation or chemotherapy

Using the techniques of complement fixation, immunofluorescence, and in vitro lymphocyte transformation (LTF), we studied the humoral antibody and cell-mediated immunity to cytomegalovirus (CMV) in normal subjects, in patients with cancer receiving localized or nonlocalized radiation, and in renal transplant recipients on immunosuppressive chemotherapy. The LTF activity was determined by the whole blood microassay, using four strains of CMV (AD-169, Davis, Veca, and Towne), AD-169 early antigen, and phytohemagglutinin (PAH). The renal transplant subjects manifested significantly depressed LTF responses to PHA and CMV and frequent presence of immunoglobulin M and early antigen-specific antibody response. The depressed LTF response to CMV recovered significantly 2 years after transplantation. The cancer patients were also characterized by a profound drop in LTF responses to PHA and CMV and in immunoglobulin M and early antigen-specific antibody response after nonlocalized radiation. LTF responses to AD-169 and Towne strains were found to be higher than those to Davis and Veca strains. The LTF response to PHA declined with age. However, LTF responses to specific CMV antigens were found to be somewhat increased with advancing age. These observations suggest that transplantation-associated suppression of CMV-specific cell-mediated immunity may improve a few years after transplantation despite continued immunosuppressive therapy.

Cell‐mediated immunity to cytomegalovirus in patients receiving immunosuppressive therapy

Employing the techniques of complement-fixation (CF), immunofluorescence (IF), and in vitro lymphocyte transformation (LTF), the humoral antibody response and cell-mediated immune (CMI) response to cytomegalovirus (CMV) were studied in the serum and peripheral blood lymphocytes in 19 normal children (controls) and 23 patients with renal disease who were receiving immunosuppressive therapy or undergoing hemodialysis. The LTF activity was determined by the whole blood microassay using two strains of CMV (AD-169 and Davis) and phytohemagglutinin (PHA). The antibody level responses to CMV in different groups of subjects were generally similar. The LTF response to PHA as evidenced by delta cpm activity was moderately depressed in immunosuppressed and hemodialyzed subjects compared to the response observed in the controls. The mean delta cpm activities in response to AD-169 and Davis strains of CMV in seropositive immunosuppressed patients were about one-fifth and one-third lower respectively than those of seropositive normal controls. These observations suggest that an impairment of CMV specific cellular immunity may be an important mechanism underlying the increased susceptibility to CMV infections in patients with chronic renal disease who receive immunosuppressive therapy.

Cell-mediated immune response to liver tissue antigen and hepatitis B surface antigen after infection with hepatitis B virus in humans.

The in vitro correlates of cell-mediated immunity to liver tissue antigens and hepatitis B surface antigen (HBsAg) were studied in groups of subjects with acute and chronic hepatitis B virus (HBV) infection and in a population of HBV-seronegative controls. The technique of in vitro lymphocyte transformation (LTF) was employed in these studies. No LTF response to liver-specific antigen and HBsAg was observed in the control population. LTF activity in response to HBsAg was present in 11 of the 14 subjects with acute type B viral hepatitis during the early phase of the disease, and eight of these subjects also had LTF reactivity to liver antigen. During the convalescent phase the LTF reactivity to these antigens usually disappeared. More than 70% of patients with chronic carriage of HBsAg who had elevated levels of liver enzymes showed LTF responses to HBsAg, and a significant number of these subjects also exhibited LTF response to liver-specific antigen. On the other hand, chronic HBsAg carriers who persistently showed normal liver enzyme values (asymptomatic carriers) failed to show significant responses to liver antigen or HBsAg. It is suggested that the persistence of cellular reactivity to liver antigens may lead to the establishment of chronic liver disease.

Hepatitis B surface antigen-specific cell-mediated immune responses in human chronic hepatitis B surface antigen carriers.

The presence of hepatitis B surface antigen (HBsAg) and antibody (anti-HBs), hepatitis B e antigen (HBeAg) and antibody (anti-HBe), the nature of T-cell function, and specific cell-mediated immunity to HBsAg were determined and evaluated serially in groups of subjects with chronic HBsAg carrier states and in seronegative controls. The techniques of in vitro lymphocyte transformation, spontaneous rosette formation, radioimmunoassay, reverse passive hemagglutination, passive hemagglutination, rheophoresis, and liver function tests were employed for these studies. For the lymphocyte transformation assay, multiple concentrations of phytohemagglutinin and purified HBsAg were used as stimulants. Cell-mediated immunity to HBsAg was detectable in 50% of the chronic HBsAg carriers (responders) at one or more concentrations of HBsAg. The remaining carriers (nonresponders) and controls failed to manifest HBsAg-specific lymphocyte transformation activity. The profile of the responders was characterized by elevated serum glutamic pyruvic transaminase levels, the presence of anti-HBe, high HBsAg titers, and the conspicuous absence of HBeAg in the serum. The nonresponders were characterized by normal serum glutamic pyruvic transaminase levels, the presence of HBeAg and anti-HBe, and lower HBsAg titers. These observations demonstrate the presence of specific cell-mediated immunity to HBsAg in chronic HBsAg carriers who manifest biochemical evidence of liver disease.

Age Related Differences in Cellular Immune Response to Vaccine Induced Rubella Infection

Employing the techniques of in-vitro lymphocyte transformation (LTF) and using Putnam strain of rubella virus as the antigen, the development of rubella specific cellular immune response was studied in different age groups of rubella seronegative normal subjects at various intervals after subcutaneous administration of HPV=77/DE5 live attenuated rubella vaccine. The rubella specific lymphocyte response in children ranging in age from two to twelve years was characterized by significant LTF activity at two months, followed by a gradual decrease. The response in adult subjects 18 to 35 years of age showed a slight delay initially in the appearance and the maximum LTF activity appeared to be 3--4 fold lower (P less than 0.01) than observed in the children. No difference was observed in the maximum antibody titers to rubella virus between these two groups of subjects. These observations suggest that the age related differences in the lymphoproliferative responses might be associated with adverse effects which are known to occur more frequently in adolescent and older patients than in childhood population after vaccine induced rubella infection.

Strain-specific local and systemic cell-mediated immune responses to cytomegalovirus in humans.

Employing the techniques of complement fixation, immunofluorescence, and in vitro lymphocyte transformation assay, the antibody and cell-mediated immunity to cytomegalovirus (CMV) were studied in the serum, peripheral blood lymphocytes, tonsillar lymphocytes, and cord blood lymphocytes. The study population consisted of 32 children undergoing tonsillectomy and adenoidectomy. In the lymphocyte transformation assay, three strains of CMV (AD-169, ADH-1-41, and Davis), herpes simplex type 1, and phytohemagglutinin were employed as antigens. Sixty-five percent of the subjects were found to have CMV-specific antibody activity. The lymphocyte transformation response to phytohemagglutinin was similar in all subjects. No CMV-specific lymphocyte transformation activity was detected in cultures of cord blood lymphocytes. Significant cell-mediated immunity was observed in the tonsillar lymphocytes of 30% (3/10) of the seronegative individuals and in the peripheral blood lymphocytes obtained from one such subject. Over 75% (16/21) of the seropositive subjects demonstrated cell-mediated immunity against one or more strains of CMV in the peripheral blood lymphocytes and tonsillar lymphocytes. In the lymphocyte transformation assay, no cross-reactivity was apparent between CMV and herpes simplex type 1. These studies demonstrate the presence of strain-specific systemic and mucosal cell-mediated immune response to CMV in humans. The frequency and distribution of lymphocyte transformation responses to the three CMV strains suggest antigenic heterogeneity of CMV.

Abnormalities of Cellular Immune Response in Arthritis Induced by Rubella Vaccination

The rubella-specific antibody, activity the levels of immunoglobulins and complement in the serum, the number of T cells, and the nature of cell-mediated immunity as measured by lymphocyte transformation (LTF) of circulating lymphocytes in response to rubella virus and phyto-hemagglutinin (PHA) were studied in 15 children who manifested arthralgia or arthritis after parenteral immunization with HPV-77 DE/5 or RA27/3 live attenuated rubella virus vaccines. Eighteen appropriately matched subjects who developed no post-vaccine complications were included as controls. The immunoglobulins, complement, and rubella antibody levels were similar in all subjects. Although significant rubella specific cell-mediated immunity was detectable in most control subjects, the activity was markedly depressed in subjects with arthritis. The LTF activity in response to PHA was approximately equal in both groups. These findings suggest a selective depression of cell-mediated immunity to rubella virus in subjects with arthritis associated with rubella vaccination.

Development and Characteristics of In-Vitro Correlates of Cellular Immunity to Rubella Virus in the Systemic and Mucosal Sites in Guinea Pigs

Abstract The development of rubella-specific cell-mediated immunity (CMI) in splenic and bronchial lymphoid cells and antibody activity in serum and bronchial washings was studied in guinea pigs after subcutaneous or intranasal inoculation with live attenuated HPV-77 DE/5, RA27/3 rubella vaccines or live Brown strain of rubella virus. The techniques of hemagglutination inhibition, in vitro lymphocyte transformation, and assay of migration inhibitory factor with rubella virus as the antigen were employed to determine antibody and CMI in the systemic and secretory sites. Subcutaneous immunization resulted in regular appearance of serum antibody response. However, no antibody was detected in bronchial washings. Intranasal administration of the virus preparation resulted in little or no antibody response in the serum or respiratory tract. The CMI response after subcutaneous administration of rubella virus was characterized by the specific induction of lymphocyte transformation and migration inhibitory factor activity in the spleen cells without any detectable response in the bronchial lymphoid cells. Intranasal immunization resulted in the appearance of migration inhibitory factor activity in the bronchial cells with little or no response in the spleen cells. The CMI responses were initially detected 2 weeks after immunization. Peak responses were often found at 4 weeks, and no specific migration inhibitory factor or lymphocyte transformation activity was detected after 6 weeks. The results of this study suggest the development of local cellular immune response in the respiratory tract after local application of rubella virus.

Lymphocyte transformation and macrophage migration inhibition by electrofocused and gel-filtered fractions of group A streptococcal filtrate

Culture filtrates of group A streptococci were fractionated either by isoelectric focusing on a sucrose gradient at pH 3–10, or by gel filtration on a G-75 Superfine Sephadex column. Some fractions induced lymphocyte transformation, others inhibition of macrophage migration, and others both. With the two types of fractionation here used the lymphocyte transformation activity was concentrated in a single peak, while the activity responsible for macrophage migration inhibition was scattered over multiple fractions. The significance of these findings is discussed.