Inorganic pyrophosphate: formation in bacterial photophosphorylation.

Inorganic pyrophosphate is identified as the major product of photophosphorylation by isolated chromatophores from Rhodospirillum rubrum in the absence of added nucleotides.

Elevated neopterin and beta 2-microglobulin levels in blood and cerebrospinal fluid occur early in HIV-1 infection.

Neopterin and beta 2-microglobulin (beta 2-M) concentrations in cerebrospinal fluid (CSF) and blood were measured in 56 individuals in various stages of HIV-1 infection. Elevated levels of neopterin as well as beta 2-M were found in the CSF of three patients with primary HIV-1 infection and also in subjects in the early stages of chronic HIV-1 infection, with the highest levels in HIV-1 isolation-positive people. There was a clear correlation between the concentrations of the two substances and the levels seemed to increase in parallel with progress of infection. A similar pattern was found in blood. Higher concentrations of neopterin and beta 2-M in CSF than in blood were found in patients with advanced dementia in particular. These findings indicate that the cellular immune system in the central nervous system (CNS) may be activated during the early stages of HIV-1 infection without concomitant overt neurological symptoms. The pathological processes in CNS and blood seem to develop in parallel rather than being restricted to one compartment.

Immunological changes in primary HIV-1 infection.

Homosexual men with symptomatic primary HIV-1 infection displayed a pronounced lymphopaenia with significantly depressed numbers of CD3+, CD4+ and CD8+ cells and B cells during the first week of illness. Subsequently, the CD8+ cell counts rose in parallel with numbers of CD3+ cells, atypical lymphocytes and activated (CD38+ and HLA-Dr+) cells to attain maximal levels about a month following onset of illness. In contrast CD4+ and B cell numbers remained low for an extended period of time. Early signs of a host response included a transient appearance of interferon-alpha in the blood and raised levels of neopterin and beta 2-microglobulin (beta 2-M). Neither CD4+/CD8+ cell ratio nor beta 2-M resumed completely normal values during a follow-up period of 2 years. These findings shed some light on pathogenetic events during early HIV-1 infection and suggest that the infection, following the acute symptomatic stage, usually enters a stage of chronic active rather than latent infection.

Septic arthritis caused by a gram-negative bacterium representing a new species related to the Bordetella-Alcaligenes complex.

A knee-joint exudate culture yielded on two occasions a gram-negative bacterium. Regular methods for speciation did not provide an identification. The infection was successfully treated with ciprofloxacin. The unknown isolate, CCUG 36768, was subjected to further investigation, including 16S rDNA sequencing, protein profiling, cellular fatty acid analysis, and various biochemical tests, in order to produce a species identification. The 1469 bp-long 16S rDNA sequence did not reveal identity with any known species sequence. CCUG 36768 clustered in a group of species, including Alcaligenes defragrans, Denitrobacter permanens, Taylorella equigenitalis, Alcaligenes faecalis, and four strains of Alcaligenes species without a specific species name. Bordetella species also showed a high degree of similarity with CCUG 36768. Protein profiling, cellular fatty acid analysis and computer-assisted analysis of biochemical profiles indicated similarity with Bordetella-Alcaligenes species, often close to B. holmesii and B. avium. API 20 NE indicated the profile of Moraxella species of poor identity. It is concluded that CCUG 36768 represents a new bacterial species of pathogenic potential in humans. It is related to the Bordetella-Alcaligenes group. Powerful new methods for speciation are available and it is recommended that unknown isolates from normally sterile sites be submitted for further analysis. Several isolates are required for the definition of new species.

Long-term effects on the immune system following local irradiation for breast cancer. Pokeweed mitogen induced immunoglobulin secretion by blood lymphocytes and serum immunoglobulin levels.

Previous studies have shown that the PWM driven immunoglobulin (Ig) secretion by blood lymphocytes in vitro is reduced shortly after local radiation therapy for breast cancer. In this investigation we have examined the long-term effects of radiation therapy (45 Gy) on this lymphocyte function. A total of 111 disease-free patients treated for operable breast cancer 5-6 and 10-11 years earlier were examined. The Ig classes studied were IgM, IgA and IgG. All patients belonged to a clinical trial where the effect of pre- or post-operative local radiation therapy was evaluated against surgery only. Significant reductions of the amount of spontaneously released IgM in vitro were observed among irradiated patients as compared to the unirradiated 5-6 and 10-11 years after treatment. Such a difference was not observed for IgA and IgG. The PWM induced Ig secretions did not differ significantly between the three patient groups. The long standing reduction of the spontaneous release of IgM by blood lymphocytes in vitro probably lacks clinical significance since the serum levels of this Ig class, as well as IgA and IgG, were similar in irradiated and unirradiated patients.

Impact of adjuvant chemotherapy on spontaneous and poke week mitogen triggered immunoglobulin secretion of blood lymphocytes in operable breast cancer.

The objective of this investigation was to examine further the influence of postoperative adjuvant treatment with cyclophosphamide, methotrexate and 5-fluorouracil (CMF) on the immune system in breast cancer patients and to explore whether such changes are related to prognosis. The 12 CMF courses which were given for a period of 1 year resulted in a progressive lymphopenia. The relative spontaneous secretions of IgA and IgG in vitro increased 4- and 2-fold respectively after the first 3 courses, whereas IgM secretion was unaffected. IgA and IgG secretions in PWM stimulated cultures were not changed, whereas there was a sharp decrease of IgM. The CMF-induced changes of Ig-secretions were similar in patients who developed recurrent disease during a 4-6 year follow-up (n = 11) and those who remained clinically disease-free (n = 14). The results are discussed in relation to the immunopotentiation which may occur following treatment with relatively low doses of chemotherapeutic agents.

Influence of adjuvant radiation therapy in breast cancer on PWM induced Ig-secretion by blood lymphocytes in vitro.

The capacity of PWM (poke-weed mitogen) to stimulate Ig-secretion by blood lymphocytes was examined before and at various times after local radiation therapy (46.0 Gy) for breast cancer. It was observed that the secretion of IgM, IgA and IgG were significantly reduced at completion of radiation therapy. Secretion of IgM was reduced to the highest relative extent (approximately 10% of the pretreatment value). Thereafter there was a recovery of the Ig-secreting capacity which, however, remained below the pretreatment level, 12-18 months after completion of radiation therapy. The capacity of PWM-stimulated blood lymphocytes to secrete specific antibodies against morbilli and herpes simplex virus was also significantly impaired after radiation therapy. The reduction of Ig-synthesis in vitro was not correlated with the increase in the ratio between numbers of monocytes and lymphocytes in peripheral blood after radiation therapy. Possible explanations for these results are discussed.

Bestatin treatment enhances the recovery of radiation induced impairments of the immunological reactivity of the blood lymphocyte population in bladder cancer patients.

Bestatin, an immunostimulating substance of microbial origin, was examined for its capacity to augment immune responses of blood lymphocytes in bladder cancer patients having received a full course of local irradiation (64 Gy). Following irradiation the patients became lymphopenic and the lymphocytes exhibited impaired mitogenic responses to phytohemagglutinin (PHA) and purified protein derivative of tuberculin (PPD) and reduced poke weed mitogen induced secretion of immunoglobulins in vitro. Patients who were randomized to receive daily oral Bestatin treatment exhibited enhanced recoveries of PHA- and PPD-responses and enhanced recovery of the IgM secreting capacity compared to irradiated patients who did not receive Bestatin. Repopulation of the blood lymphocyte population, however, was not enhanced by Bestatin treatment. It is concluded that Bestatin treatment may enhance the recovery of radiation induced functional defects of the immune system in cancer patients.

Effect of radiation therapy and in vitro x-ray exposure on lymphocyte subpopulations and their functions.

Radiation treatment of breast cancer patients (45.0 Gy) profoundly affected the peripheral blood lymphocytes. The number of these cells was markedly reduced with non-T-cells being more extensively depleted than T-cells immediately after radiation. The long-lasting lymphopenia, on the other hand, was mainly due to reduced number of T-cells. Antigen and mitogen stimulability, MLC reactivity, pokeweed (PWM)-induced immunoglobulin (Ig) production in vitro, and different cytotoxic functions decreased. Depletion of lymphocytes largely restored the radiation-depressed lymphocyte reactivity. The effects of in vitro exposure of blood lymphocytes to x-rays were similar to those seen after radiotherapy. Non-T-cells and T-cells with Fc-receptors for IgG were relatively radiosensitive. This latter observation agreed well with demonstrated increase of PWM-induced Ig synthesis after in vitro exposure to x-rays. T-suppressor cells defined by monoclonal antibodies were, however, radioresistant. The cytotoxic functions were reduced. No correlations were found between the pretreatment immunological status or the extent of radiation-induced immunological suppression, respectively, and prognosis.

T-cell helper activity and B-cell function of synovial and blood lymphocytes from patients with rheumatoid arthritis or other forms of chronic arthritis.

The helper effect of T cells on B-cell immunoglobulin (Ig) responses induced by pokeweed mitogen (PWM) or purified protein derivative of tuberculin (PPD) was studied in lymphocytes from synovial fluid (SF) and blood of nine patients with rheumatoid arthritis (RA) and eight patients with other forms of chronic arthritis. In PWM cultures the helper effect of SF T cells on Ig responses (IgG, IgM, IgA) of autologous and allogeneic blood B cells was lower than that of blood T cells (P less than 0.01). This decrease was more pronounced in patients with RA than in patients with non-RA. In PPD cultures no significant difference was found between the helper effect of SF T cells and blood T cells on the Ig responses of allogeneic blood B cells or on the IgG response of autologous blood B cells, whereas the helper effect of SF T cells on the IgM and IgA responses of autologous blood B cells was decreased. The Ig responses to PWM or PPD in cocultures of autologous blood B and T cells were not significantly different between patients and healthy controls. The PWM- and PPD-induced Ig responses of SF B cells were lower than those of blood B cells when cocultured with autologous blood T cells. SF B cells produced IgG but usually little IgM and IgA. Thus there was a dysfunction of SF B cells and of SF T cells in a PWM-driven system, but a fairly good helper function of SF T cells in a PPD-driven system.

Immunoregulatory function of T8 and T4 cells from synovial fluid and blood of patients with rheumatoid arthritis or other forms of chronic arthritis.

The suppressor effect of synovial fluid (SF) T8 cells and blood T8 cells on the pokeweed mitogen (PWM)-induced T4 cell-dependent immunoglobulin production of autologous blood B cells was studied in nine patients with chronic rheumatic diseases (six patients with rheumatoid arthritis (RA), one patient with juvenile RA, and two patients with other forms of chronic arthritis). The suppressor effect of SF T8 cells was of the same magnitude as that of equal numbers of blood T8 cells from patients and healthy controls. However, the relative number of T8 cells was higher among SF T cells than among blood T cells in several cases. Good synovial T8 cell suppression was also demonstrated in coculture experiments where SF T4 cells and B cells were used. In PPD (purified protein derivative of tuberculin)-stimulated cultures the suppressor effect of SF T8 cells as well as of blood T8 cells from patients and controls was lower than it was in PWM-stimulated cultures. In most patients SF T4 cells showed a much better PWM-induced helper function than did non-fractionated SF T cells. Thus the poor PWM induced helper effect of non fractionated synovial T cells was in some cases mainly due to the suppressor effect of T8 cells, whereas in some cases there was also a deficient helper function of synovial T4 cells.

T8 cell suppression of antigen- and mitogen-induced T4 cell dependent immunoglobulin production.

The suppressor effect of T8 cells on antigen-induced, as compared to pokeweed mitogen-induced, T4 cell dependent immunoglobulin (Ig) production by B cells of healthy subjects was studied. The antigens used were purified protein derivative of tuberculin (PPD) and tetanus toxoid (TT). The suppressor effect of T8 cells on IgG, IgM and IgA responses in co-cultures of T4 cells and B cells was significantly stronger in the pokeweed mitogen driven system than in PPD- and TT-driven cultures under the same experimental conditions.

An evaluation of delayed hypersensitivity in guinea pigs to various trichophytin preparations.

Guinea pigs immunized with mycelium from Trichophyton mentagrophytes in Freund's complete adjuvant were tested intradermally with the following trichophytin preparations: TrI extracted by ethylene glycol, TrII extracted by phenol and TrII, a commercially obtained preparation fromSächsisches Serumverk KG. In our experience, TrI was found to be superior to the other preparations. It showed signifiicant delayed skin reactions in all sensitized animals and none in the controls. Lymphocyte stimulation test with this preparation (TrI) gave positive results.

T lymphocyte subpopulations defined by monoclonal antibodies and FC receptor binding in relation to immunosuppression in vaccine-induced rubella infection.

Lymphocyte stimulation tests and determination of T lymphocyte subpopulations were undertaken before and 7-11 days after rubella vaccination of healthy subjects. The proliferatory response of blood mononuclear cells to phytohemagglutinin (PHA) and purified protein derivate was suppressed after vaccination, as was the immunoglobulin production of pokeweed mitogen (PWM) stimulated cultures. After removal of phagocytic cells the response to PHA was still suppressed after vaccination in most cases, but when the cultures were depleted of non-T lymphocytes the suppression was no longer demonstrable. The proportion of T lymphocytes with receptors for the Fc part of IgG increased after vaccination, whereas there was no significant change of the T lymphocyte subpopulations identified with monoclonal antibodies directed at the suppressor/cytotoxic or helper T cell subsets.

Radiosensitive t cell suppression in relation to the TG cell level in synovial fluid of patients with rheumatic disease.

Radiosensitive T cell suppression was studied in relation to the level of T cells with IgG-Fc receptor (TG cells) in synovial fluid and blood of nine patients with rheumatic diseases. Irradiation of T cells significantly decreased the proportion of TG cells in the blood of patients and healthy controls. The proportion of synovial TG cells was low in all patients before as well as after irradiation. Irradiation enhanced the helper effect of T cells on pokeweed-mitogen induced immunoglobulin production by allogeneic B cells. This increased helper effect was taken as an expression of decreased suppressor activity following irradiation. By this method suppressor activity was demonstrable in synovial T cells of six out of nine patients and in blood T cells of six out of eight patients and six out of eight healthy subjects. Thus the low level of synovial TG cells was in most patients not associated with a lack of radiosensitive synovial suppressor cell activity.

Changes in Borrelia burgdorferi-specific serum IgG antibody levels in patients treated for acrodermatitis chronica atrophicans.

The kinetics of Borrelia burgdorferi-specific serum IgG antibody values in 74 patients treated for acrodermatitis chronica atrophicans was analysed by means of enzyme-linked immunosorbent assay. At the last clinical control, there had been no clinical signs of active infection. The serological follow-up time ranged from 12 months to 5 1/2 years (median 2 years and 1 month). In 68 (92%) of the 74 patients, a significant decrease of the specific antibody values was found within 3 years after the initiation of therapy. In 53 (72%) of the patients, this decrease was found within 15 months. Most of the patients remained seropositive during the follow-up period. The results show that a significant decline of the levels of serum IgG antibodies to Borrelia burgdorferi can be expected in the majority of patients who do not exhibit clinical evidence of persistent infection after antibiotic treatment of acrodermatitis chronica atrophicans.

Polymerase chain reaction for detection of Borrelia burgdorferi DNA in skin lesions of early and late Lyme borreliosis.

The aim of this study was to evaluate the polymerase chain reaction (PCR) as a diagnostic tool for Lyme borreliosis on large numbers of samples from clinically well-defined cases of early and late cutaneous borreliosis. Skin biopsy specimens from patients with erythema migrans and acrodermatitis chronica atrophicans were analysed blindly together with an equal number of control biopsies. Using two different dilutions of each DNA specimen increased the number of total positives detected. All of the 76 control biopsies were PCR negative. Biopsy specimens from 18 of 26 (69%) erythema migrans lesions and from 22 of 36 (61%) acrodermatitis chronica atrophicans lesions were PCR positive. Fourteen post-therapy biopsies from patients with acrodermatitis chronica atrophicans were all negative, supporting the opinion that antibiotic therapy is successful in this chronic manifestation of Lyme borreliosis.

Blood lymphocyte population following 131I treatment for hyperthyroidism.

A number of immunological parameters have been monitored for up to 6 weeks following 131I treatment for hyperthyroidism in Graves' disease. The aim was to examine whether this isotope treatment normalizes or further accentuates some immunological abnormalities which may be a manifestation of autoimmune reactions in these patients. It was confirmed that both the cellular composition and immunological reactivities of the patients' blood lymphocytes were abnormal before treatment. After 131I administration a slight lymphopenia occurred and the ratio between CD4 and CD8 positive T lymphocytes (helper-inducer/suppressor-cytotoxic), which was increased before treatment, increased further. Moreover, PWM-triggered IgM secretion in vitro was reduced by 50%. No other immunological parameters studied, such as secretion of other Ig classes, mitogenic responses of lymphocytes, and distribution of other lymphocyte subsets, changed to any detectable extent. It remains speculative whether the 131I-induced changes of the immune system may further accelerate the underlying autoimmune disease processes.

Influence of adjuvant tamoxifen on blood lymphocytes.

The blood lymphocyte population was studied in 23 breast cancer patients treated with adjuvant tamoxifen for 1.5-2 years, and in an equal number of control patients. The size and cellular composition of the blood lymphocyte population, as assessed by monoclonal antibodies directed against various subsets, did not differ between the two patient groups. However, lymphocytes from the tamoxifen-treated patients exhibited a significantly lower NK activity against K562 cells. In contrast, the proliferative response of lymphocytes to ConA was significantly higher. These results indicate that tamoxifen may modulate the immune system.

Elevated serum beta-2-microglobulin--a prognostic marker for development of AIDS among patients with persistent generalized lymphadenopathy.

For evaluation of its prognostic value, the level of serum beta-2-microglobulin was determined in early serum samples from 88 patients with persistent generalized lymphadenopathy in a prospective longitudinal study. Patients with serum beta-2-microglobulin greater than 2.6 mg/l were found to have a significantly higher risk of developing AIDS earlier when compared to patients with a lower level (p less than 0.001).