Requirement of PI3-kinase activity for the nuclear transport of prolactin in cloned murine T lymphocytes.

The murine T-cell clone, L2, requires both IL2 and PRL to proliferate. Proliferation and selected IL2-driven gene expression are blocked by treatment with rapamycin. Since prolactin translocation to the nucleus is IL2 dependent and required for proliferation, experiments were performed to identify activation pathways that might be involved in nuclear transport and proliferation. L2 cells were stimulated with IL2 in the presence and absence of the mTOR inhibitor rapamycin, PI3-kinase inhibitors (wortmannin, LY294002), the p38 MAP kinase inhibitor SB203580 and the vitamin D analog calcipotriol. The immunosuppressant rapamycin markedly inhibited IL2-induced proliferation and prolactin translocation to the nucleus. Similarly, wortmannin and LY294002 inhibited IL2-induced proliferation and markedly decreased the amount of prolactin transported to the nucleus. SB203580 and calcipotriol partially inhibited IL2-induced proliferation but had no effect on prolactin translocation. None of the inhibitors affected Lucifer Yellow uptake indicating that rapamycin, wortmannin and LY294002 did not inhibit early endosomal formation but rather worked to inhibit prolactin translocation at a later point in the retrograde transport pathway.     

Modulation of prolactin expression in human T lymphocytes by cytokines

Besides its pivotal role in reproduction, the polypeptide hormone prolactin (PRL) has immunomodulatory properties. Whereas the bulk of circulating PRL is produced by the pituitary, PRL is also produced by the decidua, the myometrium, the mammary gland and leukocytes. Extrapituitary PRL expression is regulated differently from that in the pituitary, due to the use of an alternative promoter. Here we show for the first time that in T lymphocytes PRL expression is subject to regulation by cytokines. We established that both IL-2 and IL-4 reduced PRL mRNA levels in T lymphocytes to 25 and 28% of control values, respectively. PRL mRNA expression was inhibited to a lesser extent by IL-1beta, which decreased PRL mRNA levels to 58% of control values.     

Panic disorder and immunity: few effects on circulating lymphocytes,mitogen response,and NK cell activity

Altered immune measures are commonly found in major depression (MD), however, less is known about the immune system in anxiety disorders. We examined quantitative and functional in vitro immune measures in patients with panic disorder (PD), which is often comorbid with MD. Fourteen otherwise healthy medication-free adults (ages 23-49; 11 female) meeting SCID-UP DSM-IIIR criteria for PD with agoraphobia and without current MD, were compared with 14 subjects free of PD, MD, or other major psychiatric disorders, matched by gender, age, and racial background. PD was associated with decreased percentage (p<.03) and total (p<.03) circulating CD19+ B lymphocytes, but no differences in other enumerative lymphocyte measures. Mitogen responses (Con A, PHA, PWM) did not differ except for possibly decreased PHA in PD (p<.06). NK cell activity did not differ between PD and control subjects. The few immune measure changes in PD contrast with those found in MD, providing further evidence for the specificity of immune changes in psychiatric disorders.     

B and T lymphocytes in man. III. B, t, and "null" lymphocytes in multiple sclerosis.

T, B, and "null" lymphocytes were determined in peripheral blood obtained from 24 control subjects and 15 multiple sclerosis patients. The total number of lymphocytes and T lymphocytes were reduced significantly in multiple sclerosis patients as compared with those of controls. Although the percentage of B lymphocytes increased in these patients, the absolute number of B lymphocytes was similar to controls. The mitotic response of lymphocytes from multiple sclerosis patients to phytohemagglutinin and pokeweed mitogen in 7-day-old cultures was decreased as compared with that of controls.     

Ecto-5'nucleotidase activity in B and T lymphocytes and monocytes of patients with multiple sclerosis

Ecto-5'nucleotidase activity in the plasma membrane of B and T lymphocytes and cultured and noncultured monocytes from patients with multiple sclerosis and from age- and sex-matched control subjects was determined by radioisotopic assay. The activity on lymphocytes and noncultured monocytes was normal, but the activity on cultured monocytes was found to be higher than normal in 70% of patients with active disease and lower than normal in 85% of patients with chronic inactive disease. Immunofluorescence staining with monoclonal antibodies, carried out in parallel with each assay, did not show any phenotypic differences between each cell population and its control.     

T and B lymphocytes in the cerebrospinal fluid of various neurological diseases

Summary Cerebrospinal fluids (CSF) from 66 patients with a variety of neurological disorders were studied for total protein content, absolute amount of albumin, IgA, IgG and IgM, as well as their quotients (fraction to total protein ratio), cell numbers and B cell and T cell levels. In addition, the percentage of B cells and T cells in the blood was determined in 34 patients and serum immunoglobulin levels were estimated in 51 patients. In noninflammatory diseases of the CNS, the percentage of B cells was slightly higher and T cell levels were lower in the CSF in comparison to corresponding blood values. The B cell to T cell ratio in viral meningitis was altered in the CSF. An apparent increase in the T cell level led to a decrease of B cell values. Similar changes were also found in optic neuritis. The percentage of T cells was higher in relapsing multiple sclerosis than in the chronic progressive form. There were less striking changes in the B cell to T cell ratios in the CSF of other inflammatory diseases of the CNS.

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Zusammenfassung Von 66 Patienten wurden im Liquor außer Zellzahl, den B-und T-Zellen noch der Gesamteiweißgehalt sowie die absoluten Werte von Albumin, IgA, IgG, IgM und deren relativer Anteil am Gesamteiweiß untersucht. Im Blut war die Bestimmung der B- und T-Zellen bei 34 Patienten, die der Immunglobuline bei 51 Patienten möglich. Bei nicht entzündlichen Erkrankungen des zentralen Nervensystems liegt der prozentuelle Anteil der T-Zellen im Liquor zumeist etwas niedriger, der der B-Zellen etwas höher als die entsprechenden Blutwerte. Bei viralen Miningitiden kommt es zu einer deutlichen Verschiebung dieses B:T-Zellen-Verhältnisses, und zwar zu einem Ansteigen der T-Zellen bei daraus resultierendem B-Zellen-Abfall. Ähnliche Relationsveränderungen sind bei retrobulbären Neuritiden, die als Erstsymptome einer MS auftreten, zu finden. MS-Fälle mit schubförmiger Verschlechterung weisen einen höheren Anteil an T-Zellen auf als MS-Fälle mit chronisch progredientem Verlauf. Eitrige Meningitiden, Myelitiden und Polyneuropathie-Syndrome zeigen die Tendenz des T-Zellen-Anstieges und B-Zellen-Abfalles im Liquor in geringerer Ausprägung.

     

T and B lymphocytes in patients with Down's syndrome.

Individuals with Down's syndrome are thought to have abnormalities of immune function. Studies to quantify the number of peripheral blood T and B lymphocytes and serum immunoglobulins in 12 individuals and 12 sex and age matched control subjects were performed. Hepatitis B antigen and antihyroglobulin antibodies as markers of possible immune dysfunction were determined. The numbers of circulating T and B cells, and the level of serum immunoglobulins in children with Down's syndrome did not differ from nonretarded control children. Circulating hepatitis B antigen and antihyroglobulin antibodies were not present. These studies indicated that quantitative abnormalities of T and B cells are not present in children with Down's syndrome. The data did not exclude the existence of qualitative abnormalities.     

Distribution of somatostatin receptors on murine spleen and Peyer's patch T and B lymphocytes

Recent evidence suggests that the neuropeptide somatostatin (SOM) plays an immunoregulatory role. We demonstrated previously that SOM inhibits concanavalin A-induced cell proliferation and immunoglobulin synthesis by murine Peyer's patch and splenic lymphocytes. Available data suggest that these effects are in part mediated by specific SOM receptors expressed by lymphocytes, but as yet these receptors have not been characterized. Using cytofluorimetry we investigated the distribution and specificity of binding of fluorescent SOM (SOM*) to murine Peyer's patch and splenic T- and B-lymphocyte subpopulations. The specificity of binding was confirmed by radioassay. T and B cells from both organs showed specific binding of SOM*. In Peyer's patches, approximately 50% of all cell populations studied (whole, T- and B-cell-enriched) bound SOM specifically and this was significantly higher than the corresponding splenic lymphocyte populations. Eighty to eighty-four percent of Peyer's patch Thy1.2+, Lyt1+, or L3T4+ cells and 94% of Lyt2+ cells bound SOM. Greater than 80% of B cells from this organ bound SOM (sIgA+ = sIgM+ greater than sIgG+ cells). In spleen, approximately 30% of Thy1.2+, Lyt1+, or L3T4+ cells bound SOM and this was significantly less than the proportion of Lyt2+ cells (53%) which did so. More sIgA+ (89%) than sIgG+ (66%) than sIgM+ (55%) B cells bound SOM*. Although we have previously shown that the effect of SOM on immunoglobulin synthesis was relatively isotype-specific (IgA synthesis was predominantly affected, especially in Peyer's patches) this cannot be explained solely on the basis of preferential expression of SOM receptors by distinct lymphocyte subsets. Instead, it is probably the result of the specific immunological microenvironment in which the lymphocytes reside.     

The proliferation rate of T and B lymphocytes in cerebrospinal fluid

Summary An immunocytochemical method is presented by which individual proliferating T and B lymphocytes can be detected in the cerebrospinal fluid (CSF) of patients with inflammatory and neoplastic diseases of the central nervous system. The absolute numbers of proliferating T and B lymphocytes were approximately equal, so that only a minority of the strongly prevailing T-cell population proliferates in the CSF during the immune process. The highest proliferation (25.7%) was found within the small subset of activated, i.e. cytoplasmic immunoglobulin-containing, B lymphocytes. This provides further evidence that these cells are not the terminally differentiated plasma cells of classical cytology but progenitor cells capable of further proliferation. B-cell lymphomas could be easily identified. The detection of abnormally proliferating cell subsets could help in the differentiation of opportunistic infection from leukaemic infiltration     

The effect on plasma prolactin levels of interictal epileptiform EEG activity.

Because of the known effects of seizures on plasma prolactin, the plasma prolactin levels were measured before and after generalised interictal epileptiform activity was provoked in the EEG in five epileptic patients. The findings were compared with those obtained in five normal subjects and three epileptic patients who were also exposed to flicker stimulation, but who did not develop a photoconvulsive EEG response. There was no significant difference in baseline prolactin values, and levels did not change with photic stimulation or in response to the presence of generalised epileptiform activity in the EEG.     

Lymphocyte subsets in relapsing-remitting Multiple Sclerosis: A longitudinal study of B lymphocytes and T lymphocytes

Abstract

Changes in lymphocyte subset populations may provide dues to the dysimmune mechanisms involved in relapsing remitting multiple sclerosis (RRMS). The lymphocyte subgroup CD4+CD45RA+, thought to be responsible for the induction of suppression is decreased in some patients with MS compared to controls. A possible role for another lymphocyte subset, CD19+CD5+ lymphocytes; has been proposed in autoimmune diseases and multiple sclerosis (MS). To expand this we studied CD4+CD45RA + (T) lymphocytes and CD19+CD5+ (B) lymphocytes in nine patients with relapsing-remitting MS (RRMS) and nine controls. The patients were examined monthly for an average of ten months and nine relapses were observed in seven patients. One patient underwent monthly gadolinium enhanced magnetic resonance imaging (MRI). Normal percentages CD4+CD45RA+ lymphocytes were found in patients with RRMS. No significant abnormalities in the CD19+CD5+ lymphocyte subpopulation were noted, although a tendency for higher percentages of this subset (approaching statistical significance, P = 0.056) was detected. [Neurol Res 1994; 16: 385-388]     

The effect of prolactin on glutamate decarboxylase activity and GABA concentration in hypothalamic slices

The effect of prolactin on the activity of GABA-related enzymes and GABA concentration were studied in hypothalamic slices incubated in vitro. After short periods of incubation (up to 40 min), prolactin (0.25 μg/ml) added to the incubation medium produced a significant increase (21% at 20 min of incubation) in glutamic acid decarboxylase (GAD) activity in the hypothalamic slices. A higher concentration of prolactin (1.0 μg/ml) produced a slight but significant decrease (8% at 20 min of incubation) in hypothalamic GAD activity. However, after longer periods of incubation (over 8 hr), both doses of prolactin induced a sustained increase in hypothalmic GAD activity, a response which depends upon protein synthesis. No changes were observed in GABA-transaminase (GABA-T) activity of hypothalamic slices incubated in the presence of prolactin. Prolactin decreased GABA concentration in the hypothalami incubated for 10 hr and, at the same time, increased GABA release into the medium. These results indicate that prolactin modifies the synthesis and release of hypothalmic GABA and suggest the existence of a feedback mechanism that prolactin may exert directly at the hypothalamic level.     

Dopamine effect on the mitochondria potential in B lymphocytes of schizophrenic patients and normal controls

Brain metabolic abnormalities and aberrant dopamine (DA) metabolism have been reported in patients with schizophrenia. The authors hypothesized that mitochondria is a primary target of damage by increased free radical generation secondary to increased DA metabolism by monoamine oxidase (MAO). Epstein-Barr virus (EBV)-transformed human B-lymphocytes cell lines derived from patients with schizophrenia and normal controls were incubated in the absence or presence of DA, hydrogen peroxide (H2O2), or rotenone (Rot). The cells were then stained with rhodamine 123 (Rh 123) and analyzed for uptake using flow cytometry. Compared with untreated cells, DA significantly decreased Rh 123 uptake by the mitochondria. This effect was similar to the control cells treated with H2O2 or Rot. Nevertheless, there were no differences in Rh 123 uptake between the cells of schizophrenic patients and normal controls. This study shows that DA can impair the mitochondria membrane potential but that mechanism may not be evident in schizophrenia.     

Bradykinin B1 receptor expression and function on T lymphocytes in active multiple sclerosis

BACKGROUND: Lesion development in MS is initiated by migration of inflammatory cells into the central nervous system, a process dependent on endothelial cell-lymphocyte interaction. Bradykinin B1 receptor is a membrane-bound G protein-coupled receptor shown to be upregulated on the surface of various cells types during inflammation. OBJECTIVE: To assess the expression and function of the bradykinin B1 receptor on T lymphocytes from MS patients. METHODS: The authors used multiplex polymerase chain reaction amplification and Western blot techniques to demonstrate B1 receptor expression by T cells. A modified Boyden chamber assay also was used to assess the effect of B1 agonist and antagonist on T cell migration. RESULTS: The authors demonstrated that the expression of B1 receptor was upregulated on T cells derived from peripheral blood of MS patients. Expression of this receptor was upregulated on T cells from patients with secondary progressive MS and relapsing-remitting patients in active relapse. Expression was lower in relapsing remitting patients in remission and least in control subjects, including patients with epilepsy, chronic inflammatory demyelinating polyneuritis, and systemic lupus erythematosus. In vitro treatment of cells from healthy control subjects with tumor necrosis factor-alpha and interferon-gamma also induced the expression of B1 receptors. The authors also found that the significantly higher rate of migration of MS T lymphocytes, compared with control subjects in the Boyden chamber assay, could be prevented by the addition of the selective and stable B1 agonist Sar (D-Phe8) desArg9-BK. CONCLUSION: The authors demonstrate that B1 receptors are upregulated by T lymphocytes during the course of MS and that signaling through this receptor with a B1 agonist can negatively regulate T-cell migration in vitro.