Search PubMed⌕ Search

Biomedical subjects

R Pabst

Publications and source records attributed to R Pabst.

At least 73 records · Page 4Linked to original sources
PubMed1999

Quantification of B, T and null lymphocyte subpopulations in the blood and lymphoid organs of the pig.

Research on the pig's immune system is not only of general biological interest; the pig is also becoming more important as a large animal model in human biomedical research, e.g. as a donor for xeno-transplantation. With the increasing panel of monoclonal antibodies against porcine lymphocyte markers it is possible to gain more insight into the distribution and phenotype of lymphocyte subpopulations in the pig. In this study we investigated B cells (surface IgG: sIgG, sIgM and sIgA) and T cells (CD2, CD4, CD8, 8/1, MAC320) in the peripheral blood (pBL), thymus, spleen, tonsil, mesenteric and inguinal lymph nodes (mLN, iLN), jejunal and ileal Peyer's patches (jejPP, ilPP) in Göttingen minipigs. A flow cytometric technique was employed which enabled three color indirect immunofluorescence. B cell stained for surface IgG and surface IgA were found only in small percentages. Surface IgM positive cells were distributed at higher rates, with up to 24.9% in the iLN. Up to 64.2% of CD4+ and up to 73.1% of CD8+ cells were observed in the thymus. Most of the CD4+ cells were CD4/CD8 double positive cells. These cells were mostly triple positive in combination with CD2. A larger fraction of CD2- were CD8- which are taken to be NK cells. MAC320, a marker for a subtype of gamma/delta T cells, was predominantly found on cells in the pBL. The standardized flow cytometric technique produced comparable data on the distribution of major lymphocyte subpopulations in the blood and different lymphoid organs of the pig. The results provide a basis for future studies using the pig as animal model.

Animals↗
PubMed1999

Quantification of proliferating lymphocyte subsets appearing in the intestinal lymph and the blood.

Lymphocyte emigration from the intestinal wall via lymphatics is necessary to maintain gastrointestinal immunity and also to connect the different parts of the mucosal immune system. In the present study the numbers and time kinetics of proliferating lymphocyte subsets leaving the gut wall via intestinal lymphatics were analysed in mesenteric lymph node adenectomized minipigs (n = 8). After cannulation of the major intestinal lymph duct, afferent lymph was collected under non-restraining conditions. In four pigs lymphocytes taken from the intestinal lymph and blood were incubated in vitro with the thymidine analogue bromodesoxyuridine (BrdU) to label all lymphocytes in the S-phase of the cell cycle. The other four pigs received a single i.v. injection of BrdU 1 week after cannulation. The initial percentage of BrdU+ lymphocyte subsets in the intestinal lymph 15 min after BrdU injection was comparable to that after the in vitro labelling (1.5 +/- 0.7% in T cells, 10.6 +/- 1.6% in IgM+ cells and 30.0 +/- 11.9% in IgA+ cells). From this level onwards, the percentage of in vivo labelled BrdU+ lymphocyte subsets reached a maximum at 12 h after BrdU application. A different pattern of BrdU+ subsets was seen in the blood. After an early peak at around 3-4 h, the frequency of BrdU in vivo labelled cells decreased. Each subset had a maximum between 12 h and 48 h after BrdU application (maximum of BrdU+ CD2+ T cells at 12 h, 4.6 +/- 1.5%; IgM+ BrdU+ at 48 h, 8.8 +/- 3.3%). The present results provide a basis to determine the time necessary for induction of specific intestinal immunity during oral vaccination studies.

Animals↗
PubMed1999

The influence of age and breeding conditions on the number and proliferation of intraepithelial lymphocytes in pigs.

The number and subset composition of intraepithelial lymphocytes (IEL) were studied in pigs in different age groups ranging from 1 day to 4.5 years. There were no major differences between the numbers of IEL in the jejunum and ileum. The postnatal increase of IEL largely depended on the breeding conditions: in germ-free animals there was a constant level, while in conventionally bred pigs the numbers increased more obviously than in specified pathogen-free (SPF) pigs. As the numbers of IEL can rise due to increased influx from other organs, decreased emigration, decreased apoptosis or local proliferation, the incorporation of the DNA precursor bromodeoxyuridine (BrdU) by IEL was studied after different labelling protocols. There were low but definite numbers of BrdU+ IEL 1 h after BrdU, indicating in-situ proliferation in conventional animals with a tendency to a higher index in the jejunum than the ileum. Repetitive labelling protocols for 14 days resulted in 12-20% BrdU+ IEL, which may be caused by local proliferation and immigration of lymphocytes produced in other lymphoid organs as documented for the pig. Future studies need to focus on the factors regulating local proliferation and the migration of IEL in different species.

Aging↗
PubMed1999

Increased expression of activation markers and adhesion molecules on lung T-cells compared with blood in the normal rat.

Lymphocytes play an important role in many lung diseases and are routinely accessible by bronchoalveolar lavage (BAL). Lymphocytes from the BAL (BAL pool) have a different subset composition to those from peripheral blood, consisting mainly of activated T-cells. The aim of this study was to examine whether preferential migration of activated T-cells to the bronchoalveolar space or factors of the specific microenvironment mediate this phenomenon. The expression of adhesion molecules and cellular activation markers (intercellular adhesion molecule-1, leukocyte function-associated antigen-1, CD2, CD44, interleukin-2 receptor and L-selectin) was studied on T- and B-cells not only in the BAL and peripheral blood (blood pool), but also in the compartments in between, such as the lung vascular perfusate (marginal pool) and the lung interstitium (interstitial pool), with the experiments being performed simultaneously in the same animals. Low levels of adhesion molecule expression were observed on T-cells in the blood and marginal pool, medium levels in the lung interstitium and the highest levels in the BAL. "Memory" (CD45R(low)) and "naive" (CD45R(high)) T-cells in the lung compartments showed a higher expression of adhesion molecules compared with blood. However, the predominating CD45R(low) T-cells showed a significantly higher expression than the CD45R(high) cells, indicating that CD4+ CD45R(high) T-cells had changed their phenotype to CD45R(low). In conclusion, a high level of expression of leukocyte function associated antigen-1 and intracellular adhesion molecule-1 on the bronchoalveolar lavage and interstitial T-cells is more likely to be the result of local, lung-specific induction than a prerequisite for migration into the bronchoalveolar space.

Animals↗
PubMed1999

Acute laryngitis in the rat induced by Moraxella catarrhalis and Bordetella pertussis: number of neutrophils, dendritic cells, and T and B lymphocytes accumulating during infection in the laryngeal mucosa strongly differs in adjacent locations.

Infectious laryngotracheitis results in fulminant respiratory distress. During the disease, the subglottic mucosa is selectively infected and swollen, the reason for this preference being unknown. Therefore, in the present study the immunoreaction of the laryngeal mucosa was studied in the rat after inhalation of either heat-killed Moraxella catarrhalis (PVG rats) or application of viable Bordetella pertussis (BN rats). The number of neutrophils, macrophages, dendritic cells, and T and B lymphocytes was determined in the mucosa of the supraglottic, glottic, and subglottic area of the larynx as well as in the trachea. After application of the pathogens, the mucosa of the subglottic area was significantly more affected than the glottic mucosa. Already 1 h after application of M. catarrhalis, not only neutrophils but also dendritic cells and T and B lymphocytes were found both subepithelially and within the epithelium. They showed a similar kinetic progression, although at a different level. Two hours after application of M. catarrhalis, at the peak of inflammation, dendritic cells (173 +/- 10 cells/0.1 mm2) outnumbered neutrophils (54 +/- 9 cells/0.1 mm2), T lymphocytes (25 +/- 2 cells/0.1 mm2), and B lymphocytes (4.3 cells/0.1 mm2). The subglottic area (and the trachea) contained about three to five times more cells than the glottic area. In contrast, the number of local macrophages was lower in the subglottic area (24 +/- 5 cells/0.1 mm2) compared with that of the glottic area (38 +/- 6 cells/0.1 mm2), and did not change after application of both M. catarrhalis and B. pertussis. Thus, infectious laryngotracheitis in the rat closely resembles the clinical picture in children. In addition, the present results show a major difference in cellular influx in the mucosa of the glottic and subglottic area. This demonstrates that even in two closely adjacent locations, inflammatory responses of different magnitudes can occur, and it underlines the importance of regulatory mechanisms specific for the respective microenvironment.

Animals↗
PubMed1999

Immunohistological characterization of leukocytes in the lungs of healthy mice and after bacterial intratracheal infection.

Leukocytes in the peripheral lung parenchyma of mice have not been characterized histologically during bacterial infection. The aim of this study was to investigate (a) the immunohistological characteristics of healthy murine lungs and (b) the cell kinetics during acute inflammation. BALB/c and MF1 mice were examined; as well as transgenic mice with the gene defect of cystic fibrosis (CF) in the airways as an animal model for this disease. MF1 mice served as controls for the transgenic animals. Lavaged and perfused lungs were snap frozen. B and T lymphocytes, CD4+ and CD8+ cells, dendritic cells, neutrophils and a subset of macrophages were enumerated on cryostat lung sections. The lung tissue and bronchoalveolar lavage (BAL) of BALB/c mice, infected intratracheally with Haemophilus influenzae type b (Hib), were studied at different time points after infection. In the lungs of healthy mice, including CF mice, the largest population was that of T cells, CD4+ cells being always more frequent than CD8+ cells. During acute inflammation the number of neutrophils in the lung parenchyma and BAL increased strongly within the first hours after bacterial instillation and reached baseline levels within one week. This study provides a semi-quantitative analysis of immunocompetent cells in normal and infected murine lung tissue. Differences in cell numbers are found between different strains. Moreover, the cellular reaction during Hib infection in mouse lungs is dominated by neutrophils, as expected in a primary immune response. In uninfected CF mice the numbers and distribution of immune cells in the lung tissue are normal, indicating that the cellular defense is adequate.

Animals↗
PubMed1999

Lymphocyte dynamics in the pulmonary microenvironment: implications for the pathophysiology of pulmonary sarcoidosis.

It is generally accepted that lymphocytes play a role in sarcoidosis. Lymphocyte numbers and in particular certain subsets are increased in the bronchoalveolar lavage fluid, and these parameters have been used as indicators for prognosis. To understand the pathophysiology of sarcoidosis the localisation and kinetics of lymphocytes in the normal lung have to be known. Lymphocytes are found in different compartments of the lung: the pulmonary vascular bed with the marginal lymphocyte pool, intraepithelial lymphocytes, the lamina propria of the bronchial tree, at a young age and under certain pathological conditions the bronchus-associated lymphoid tissue, the interstitial lymphocyte pool and the lymphocytes in the bronchoalveolar space as recovered by bronchoalveolar lavage. The lymphocyte subsets differ in these compartments. The number and subset composition are influenced by the balance of immigration, regulated by adhesion molecules, local proliferation, apoptosis and migration. In sarcoidosis lymphocyte proliferation and cell death in the bronchoalveolar space are increased several fold in the lung. More studies on regulatory factors of lymphocyte kinetics are needed in the lung of sarcoidosis patients before new therapeutic strategies can be tested.

Apoptosis↗
PubMed1998

Centrally applied NPY mimics immunoactivation induced by non-analgesic doses of met-enkephalin.

Neuropeptide Y (NPY) and endogenous opioids (EOPs) such as methionine-enkephalin (Met-enk) regulate similar physiological responses, but it is not known whether nociceptive and immune responses also show analogy after intracerebroventricular (i.c.v.) application. Dose-response studies show that Met-enk stimulates the blood granulocyte and splenic natural killer (NK) cell function of Lewis rats at a low dose (10(2) ng/kg, i.c.v.), whereas a high dose (10(5) ng/kg) causes suppression of innate immune functions associated with analgesia in the hot-plate test. At 15 min, 1 h and 24 h after i.c.v. application, both Met-enk (10(2) ng/kg) and NPY (1 ng/kg) produced similar effects: An initial suppression of innate immune function was followed by a long lasting stimulatory action on cell functions and serum interleukin-6 (sIL-6) levels. Thus, central NPY application resembles Met-enk-induced immunostimulation at doses not affecting nociception, suggesting an involvement of both peptides in shaping stress-induced immunomodulation of the non-analgetic form, possibly via activation of a common immunomodulatory effector mechanism.

Adjuvants, Immunologic↗
PubMed1998

Brain NPY Y1 receptors rapidly mediate the behavioral response to novelty and a compartment-specific modulation of granulocyte function in blood and spleen.

Neuropeptide Y (NPY) alters behavioral activity and innate immune functions of rats within minutes of intracerebroventricular (i.c.v.) application. Using combinations of the Y1-5a,b(6) agonist NPY, the Y1,3,5 agonist [Leu31-Pro34]NPY (LP-NPY), and the selective Y1 antagonist BIBP3226 (BIBP), we investigated whether the NPY-Y1 receptor (Y1R) subtype regulates NPY-induced behavioral and immunological effects at 15 min after i.c.v. application. Administration of both NPY and LP-NPY decreased rearing activity in the open field and suppressed granulocyte function in the blood. These effects were blocked by BIBP pre-treatment. In contrast to the blood, NPY and BIBP+NPY treatments stimulated granulocyte function within the splenic compartment. In addition, a blood leukophilia composed of granulocytes and NK cells was induced by NPY only. We conclude that the tested early effects of NPY are mediated by either the Y1R (rearing, blood granulocyte function), or a non-Y1R (splenic granulocyte function), or by a combined receptor activation (leukocyte mobilization). Furthermore, the immunological effects of NPY demonstrate compartment specificity.

Animals↗
PubMed1998

[How do medical students judge preclinical studies? Results of questionnaire analysis after the 1st and 2d year of studies].

BACKGROUND AND OBJECTIVE: For some years the evaluation of medical teaching has been demanded. In addition to questioning individual teachers, comparisons have to be made, but only a high response rate will provide representative results. A questionnaire study of medical students at the Hannover School of Medicine was undertaken to assess attendance at lectures, reasons for absence, importance of courses and practicals for passing preclinical examinations and as preparation for clinical studies. METHODS: Questionnaires were given to two groups of students in the summer term of 1996, a) after the first year and b) after concluding the preclinical course. RESULTS: The response rate for the two years was 92 and 87%, respectively. The reasons given for a high absentee rate at some lectures were, first of all, lack of time especially in the natural sciences--physics, chemistry and biology--but also in medical psychology, medical sociology and terminology, and secondly their lack of relevance to the clinical course. However, especially high acceptance and clinical relevance was assigned to gross anatomy. A high percentage of students were also in employment at the same time, mostly in medicine-related jobs (87%). Most students wanted continuous assessment in their compulsory subjects. CONCLUSIONS: Regular and representative questioning of students should provide encouragement for improving the medical course and a basis for ideas on reform.

Adult↗
PubMed1998

Regeneration of implanted splenic tissue in the rat: re-innervation is host age-dependent and necessary for tissue development.

The loss of spleen may lead to fatal bacterial infections. To prevent this, splenic autotransplantation has been performed in humans and experimental animals. However, there is still controversy about the protective function of this procedure. Since innervation plays an important role in splenic function, we investigated whether splenic regenerates are re-innervated, and whether this depends on the donor and host age. Splenic tissue (30 mg) was implanted into the greater omentum of either young (2 days) or old (12 months) rats, from either young or old syngeneic animals. After 3 months of regeneration, the weight of the regenerates was determined, PGP+ nerve fibers were revealed by immunohistology, and subdivided into nerve fibers of sympathetic (TH+, NPY+) or sensory (SP+, CGRP+) origin. In addition, proliferating (Ki-67 proliferation antigen+) and apoptotic cells (TUNEL technique+) were likewise investigated. No innervation of splenic regenerates was observed after implantation into old hosts, correlating with poorly developed splenic compartments. In contrast, almost normal re-innervation occurred in young hosts after implantation of both young and old splenic tissue. These regenerates showed well-developed splenic compartments and a normal number and tissue distribution of proliferating and apoptotic cells. However, after the implantation of young tissue, the final size of splenic regenerates was three times larger (140 +/- 30 vs. 40 +/- 10 mg). Thus, re-innervation of splenic implants is necessary for their subsequent development. It is determined by host age, whereas the final size of the splenic regenerates is regulated by donor age-dependent factors. This model is useful for studying both the process leading to initial innervation and the consequences of this innervation.

Aging↗
PubMed1998

[The path to "Doctor of Medicine"--how do doctoral candidates evaluate their dissertation?].

BACKGROUND AND OBJECTIVE: The value of medical dissertations has for some time been controversial. Critical opinions have, on the one hand, been directed against the effort required, its duration and the concept behind these dissertations, while asserting, on the other hand, that such dissertations hinder studying and lead to unnecessary lengthening of the medical curriculum. PARTICIPANTS AND METHOD: In collaboration with the Medical School in Hanover a questionnaire was sent to all the 189 dissertationists of one year, to be returned anonymously. It contained 16 questions about the dissertation itself, supervision. time spent on it, total duration of medical course and effect of the dissertation on the individual's medical studies. The answers were evaluated statistically by explorative data analysis. RESULTS: 181 questionnaires could be evaluated (96%). Average age of those questioned was 30.5 years, median duration of medical studies to-data was 13 semesters. For 80% it was the first attempted dissertation. 22% had been working on an entirely experimental topic (laboratory or animal experiments). The median time from starting to submitting the dissertation was 4 years, the median time of the experimental phase one year. Two-thirds of students said that work on the dissertation had not prolonged their studies. Supervision during various phases of the dissertation was overwhelmingly judged to have been "very good" or "good". Altogether 90% of dissertationists thought that it had been personally meaningful, and they would recommend it to younger costudents. CONCLUSIONS: The results of this study show that a universal criticism of medical dissertations does not stand up to detailed enquiry. Successful dissertationists highly rated their value both to themselves and to scientific medicine. The medical dissertation should therefore firmly remain as part of medical studies and of research within them.

Academic Dissertations as Topic↗
PubMed1998

Modulation of innate immune functions by intracerebroventricularly applied neuropeptide Y: dose and time dependent effects.

Centrally applied neuropeptide Y (NPY) interacts with the autonomic nervous system and the hypothalamo-pituitary-adrenal (HPA) axis activity. Since these physiological systems have been shown to modulate innate immune functions, the effects of intracerebroventricular (i.c.v.) NPY administration on leukocyte subsets in the blood, spleen and intravascular pool of the lung, blood granulocyte chemiluminescence response, and splenic natural killer (NK) cell-mediated lysis were studied in Lewis rats. Concentration-dependent NPY effects were tested at 15 min and 24 h post i.c.v. injection at dosages of 10(-6) M, 10(-9) M, and 10(-12) M. Time dependent effects were investigated at 15 min, 1 h and 24 h after i.c.v. administration of 10(-9) M NPY. Compared to saline controls, an increased number of granulocytes and NK cells in the blood, associated with a decreased granulocyte function and NK cytotoxicity was observed 15 min following NPY infusion. This initial immunosuppression was followed by long lasting stimulatory effects of NPY on the functional capacity of both cell populations when tested at 1 h and 24 h. The dosage of i.c.v. 10(-6) M NPY produced no changes, whilst 10(-9) M produced maximal, and 10(-12) M still significant effects. Results provide evidence that centrally applied NPY influences innate immunity in a dose and time dependent fashion. Cell mobilization from the vascular marginal pool is likely to be an underlying mechanism for the initial immunosuppression.

Animals↗
PubMed1998

Lymphocyte subsets in distinct lung compartments show a different ability to produce interferon-gamma (IFN-gamma) during a pulmonary immune response.

Lymphocytes play an important immunoregulatory role in pulmonary immune responses. By releasing cytokines they can control the cell-cell communication of other participating cells. Although it is well established that the lung lymphocytes, localized in distinct compartments, differ in their subset composition, little is known about cytokine production in these compartments during immune responses. Lewis rats were immunized by intravenous administration of sheep erythrocytes on day 0 and day 7 and challenged intratracheally with sheep erythrocytes on day 10. Four days after intratracheal (i.t.) challenge the composition of lymphocyte subsets (CD2+, CD4+, CD8+, B cells, natural killer (NK) cells) in the spleen, blood, lung perfusate, lung tissue and bronchoalveolar lavage fluid (BALF) was characterized, and intracellular IFN-gamma was detected in these subsets by flow cytometry. Comparing control and immunized animals, no changes were found in lymphocyte numbers, subsets or the percentage of IFN-gamma-producing lymphocytes in the spleen, blood and lung perfusate. In lung tissue and BALF, however, the absolute number of all lymphocyte subsets and the percentage of IFN-gamma-producing lymphocytes were increased. When the lymphocyte subsets were analysed an increased percentage of IFN-gamma-producing T cells was found in lung tissue (4.5 +/- 0.6% versus 12.8 +/- 1.1%) and in BALF (7.8 +/- 1.4% versus 14.8 +/- 1.9%) of immunized animals opposed to controls, this increase being seen in both CD4+ and CD8+ cells. Thus, there is an accumulation of T cells with an increased potential to produce IFN-gamma in the lung interstitium and the bronchoalveolar space during pulmonary immune responses.

Animals↗