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1.
J Exp Med ; 169(1): 321-6, 1989 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-2462611

RESUMO

Injection of rabbits with a human T cell line infected with HIV-1 caused seroconversion within 6 wk, and HIV-1 could be isolated from PBL cultures of infected rabbits. Identity of the isolated virus with HIV-1 was shown by analysis of products amplified by the polymerase chain reaction. HIV-1 infection was seen in rabbits injected with HIV-1-infected cells alone as well as in those that were first infected with HTLV-1 and subsequently with HIV-1. There were no consistent signs of disease in the rabbits infected with HIV-1 alone but HTLV-1/HIV-1-infected rabbits showed signs of illness including diarrhea and weight loss, transient neurologic impairment and, in one animal, a rapidly progressing mammary adenocarcinoma. Examination of organs taken at necropsy from both HIV-1- and HTLV-1/HIV-1-infected animals showed splenic hyperplasia and lymphocyte infiltration of the lungs, as well as moderate damage to liver and kidney in some cases.


Assuntos
Síndrome da Imunodeficiência Adquirida/fisiopatologia , Animais , DNA Viral/análise , Modelos Animais de Doenças , Soropositividade para HIV/fisiopatologia , DNA Polimerase Dirigida por RNA/metabolismo , Coelhos , Fatores de Tempo
2.
J Leukoc Biol ; 40(2): 169-81, 1986 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-3488359

RESUMO

Continuous rabbit macrophage-like cell lines were established after in vitro infection of spleen cells with either Simian virus 40, lymphotropic papovavirus or herpesvirus sylvilagus. These cell lines are characterized as morphologically similar to mature macrophages, esterase positive, and have unrearranged immunoglobulin genes. They possess macrophage functionality because they are highly phagocytic and are able to mediate an antibody dependent cell mediated cytotoxicity assay on chicken red blood cells. Northern blot analysis of total cellular RNA with a DQ alpha probe indicates that the cell lines constitutively express message for class II gene products. In addition, cell sorter analysis indicates that two of these lines display class II antigen at the cell surface.


Assuntos
Antígenos de Histocompatibilidade Classe II/imunologia , Macrófagos/imunologia , Coelhos/imunologia , Fosfatase Alcalina/metabolismo , Animais , Anticorpos Monoclonais , Antígenos de Superfície/análise , Linfócitos B/imunologia , Carboxilesterase , Hidrolases de Éster Carboxílico/metabolismo , Linhagem Celular , Transformação Celular Viral , Esterases/metabolismo , Citometria de Fluxo , Genes , Antígenos de Histocompatibilidade Classe II/genética , Macrófagos/citologia , Fagocitose , RNA Mensageiro/genética , Linfócitos T/imunologia
3.
AIDS Res Hum Retroviruses ; 9(5): 445-53, 1993 May.
Artigo em Inglês | MEDLINE | ID: mdl-8318271

RESUMO

HCN-1A is a human cerebral cortical neuronal cell line having properties consistent with cells of immature neuronal origin. This article details evidence for productive low-level infection of HCN-1A cells with human immunodeficiency virus type 1 (HIV-1). In vitro exposure to HCN-1A monolayers to a high titer of either LAV/HTLV-IIIB or HTLV-IIIMN resulted in HIV-1 p24 antigen production and a moderate increase in reverse transcriptase activity in cell-free supernatants. The cells in both LAV/HTLV-IIIB- and HTLV-IIIMN-infected cultures were passaged and proliferated as long as 5 weeks while continuing to express low levels of viral antigen. Virus-positive cells were detected by indirect immunofluorescence, using serum from an individual with acquired immune deficiency syndrome (AIDS) as well as with a gp120 monoclonal antibody. Confirmation of HCN-1A infection was provided by polymerase chain reaction analyses of both nuclear and cytoplasmic DNA and by de novo synthesis of viral proteins as shown by metabolic labeling and immunoprecipitation. Virus in cell-free supernatants from infected HCN-1A cultures was passaged to a permissive human T cell line (A3.01). HCN-1A cells had no detectable surface CD4 protein or CD4 message. However, the cells expressed the membrane glycolipids, galactocerebroside and sulfatide, possible receptors for gp120 on cells of neuronal origin. Undifferentiated HCN-1A cells provide an in vitro model for investigating potential interactions of HIV-1 with a homogeneous population of immature cortical neurons.


Assuntos
Complexo AIDS Demência/etiologia , HIV-1/patogenicidade , Neurônios/microbiologia , Sequência de Bases , Diferenciação Celular , Linhagem Celular , Córtex Cerebral/microbiologia , Córtex Cerebral/patologia , DNA Viral/genética , DNA Viral/isolamento & purificação , Feminino , Infecções por HIV/transmissão , HIV-1/genética , HIV-1/fisiologia , Humanos , Troca Materno-Fetal , Modelos Neurológicos , Dados de Sequência Molecular , Neurônios/patologia , Reação em Cadeia da Polimerase , Gravidez , Complicações Infecciosas na Gravidez/microbiologia
4.
AIDS Res Hum Retroviruses ; 7(1): 73-81, 1991 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-1707643

RESUMO

Culture supernatants from the rabbit macrophage cell line 6083 infected with a retrovirus, human immunodeficiency virus type 1 (HIV-1), were negative for reverse transcriptase (RT) expression although the line was shown to be productively infected by all other criteria tested. Supernatants from uninfected cultures of 6083, the human monocyte line U937, and from freshly isolated peripheral human monocytes, were found to contain a monocyte-derived inhibitory factor (MDIF) which interferes with a standard assay for RT. MDIF is a heat-labile activity of approximately of 40 kD. Both substrates and products of the reverse transcriptase assay are degraded by MDIF which is not affected by reduction and alkylation of disulfide bonds. MDIF is inhibited by the addition of a particular thioated oligonucleotide (S-dG30) to the reaction mixture but this addition also inhibits RT. The optimum method to minimize MDIF interference in the RT assay is by addition of ethylene glycol bis-(beta-aminoethyl ether)N,N,N',N'-tetraacetic acid (EGTA); MDIF requires divalent cations for activity and has a strong preference for calcium which is preferentially chelated by EGTA. The potential presence of this inhibitory activity should be considered when using RT levels as a measure of retroviral infection.


Assuntos
Fatores Biológicos/metabolismo , HIV-1/enzimologia , Macrófagos/microbiologia , Inibidores da Transcriptase Reversa , Animais , Cátions , Linhagem Celular Transformada , Desoxirribonucleases/metabolismo , HIV-1/fisiologia , Humanos , Macrófagos/metabolismo , DNA Polimerase Dirigida por RNA/metabolismo , Coelhos , Replicação Viral
5.
Psychopharmacology (Berl) ; 130(3): 235-41, 1997 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-9151357

RESUMO

Phorbol 12-myristate-13-acetate (PMA), a protein kinase C (PKC) activator, elevated basal cyclic AMP levels and enhanced isoproterenol-, prostaglandin E1- (PGE1), forskolin- and cholera toxin-stimulated cyclic AMP accumulation in Epstein-Barr virus (EBV)-transformed human B-lymphocytes. Staurosporine, a PKC inhibitor, significantly antagonized the increase in cyclic AMP accumulation produced by PMA, whereas the inactive phorbol ester, 4 alpha-phorbol 12,13-didecanoate (4 alpha PDD), had no effect. Basal levels of cyclic AMP and the accumulation of cyclic AMP produced by PMA, isoproterenol, PGE1, cholera toxin and the combination of these compounds with PMA were not significantly different between schizophrenics and controls. The cyclic AMP response to forskolin in the presence and absence of PMA was significantly greater in EBV-transformed human B-lymphocytes from schizophrenics. These results suggest that activation of adenylyl cyclase by forskolin is elevated in EBV-transformed B-lymphocytes derived from schizophrenics and that this elevation is further enhanced through a PKC-dependent phosphorylation mechanism.


Assuntos
Linfócitos B/efeitos dos fármacos , Transformação Celular Viral/efeitos dos fármacos , Colforsina/farmacologia , AMP Cíclico/biossíntese , Esquizofrenia/metabolismo , Adulto , Alprostadil/farmacologia , Linfócitos B/metabolismo , Toxina da Cólera/farmacologia , Feminino , Herpesvirus Humano 4 , Humanos , Isoproterenol/farmacologia , Ativação Linfocitária/efeitos dos fármacos , Masculino , Acetato de Tetradecanoilforbol/farmacologia
6.
Schizophr Res ; 5(3): 243-7, 1991 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-1662066

RESUMO

Retroviral infection has been proposed as an etiologic factor in schizophrenia. In an effort to unmask a latent retrovirus, short term cultures of peripheral lymphocytes from 15 schizophrenic subjects and nine normal controls were exposed to ionizing radiation and co-cultured with indicator cells. Reverse transcriptase activity, a marker of retroviral infection, could not be detected in any of the cultures. Our findings are further evidence against a role for retroviral infection in the etiology of schizophrenia.


Assuntos
Transtornos Neurocognitivos/microbiologia , Infecções por Retroviridae/microbiologia , Esquizofrenia/microbiologia , Psicologia do Esquizofrênico , Adulto , Linhagem Celular , Feminino , Humanos , Linfócitos/microbiologia , Masculino , Pessoa de Meia-Idade , Retroviridae/isolamento & purificação
7.
Cell Transplant ; 6(3): 231-8, 1997.
Artigo em Inglês | MEDLINE | ID: mdl-9171156

RESUMO

In vitro growth properties of three CNS-derived cell lines were compared under a variety of culture conditions. The M213-20 and J30a cell lines were each derived from embryonic CNS culture with the temperature-sensitive (ts) allele of SV40 large T antigen, tsA58, while the A7 cell line was immortalized using wild-type SV40 large T antigen. Cells immortalized with tsA58 SV40 large T proliferate at the permissive temperature, 33 degrees C, while growth is expected to be suppressed at the nonpermissive temperature, 39.5 degrees C. Both the M213-20 and J30a cell lines were capable of proliferating at 39.5 degrees C continuously for up to 6 mo. All three cell lines showed no appreciable differences in growth rates related to temperature over a 7-day period in either serum-containing or defined serum-free media. The percentage of cells in S-phase of the cell cycle did not decrease or was elevated at 39.5 degrees C for all three cell lines. After 3 wk at 39.5 degrees C, the three cell lines also showed positive immunostaining using two monoclonal antibodies reacting with different epitopes of SV40 large T antigen. Double strand DNA sequence analyses of a 300 base pair (bp) fragment of the large T gene from each cell line, which included the ts locus, revealed mutations in both the J30a and M213-20 cell lines. The J30a cell line ts mutation had reverted to wild type, and two additional loci with bp substitutions with predicted amino acid changes were also found. While the ts mutation of the M213-20 cells was retained, an additional bp substitution with a predicted amino acid change was found. The A7 cell line sequence was identical to the reference wild-type sequence. These findings suggest that (a) nucleic acid sequences in the temperature-sensitive region of the tsA58 allele of SV40 large T are not necessarily stable, and (b) temperature sensitivity of cell lines immortalized with tsA58 is not necessarily retained.


Assuntos
Antígenos Transformantes de Poliomavirus/genética , Corpo Estriado/citologia , Neurônios/citologia , Alelos , Animais , Antígenos Transformantes de Poliomavirus/análise , Sequência de Bases , Proteínas Sanguíneas/farmacologia , Ciclo Celular/fisiologia , Divisão Celular/fisiologia , Linhagem Celular Transformada/química , Linhagem Celular Transformada/citologia , Linhagem Celular Transformada/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Meios de Cultura Livres de Soro/farmacologia , Feto/citologia , Citometria de Fluxo , Imuno-Histoquímica , Dados de Sequência Molecular , Neurônios/química , Neurônios/efeitos dos fármacos , Mutação Puntual , Ratos , Fase S/fisiologia , Análise de Sequência de DNA , Temperatura
8.
Biosens Bioelectron ; 14(10-11): 751-9, 2000 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-10945449

RESUMO

Antibodies are critical reagents used in several biodetection platforms for the identification of biological agents. Recent advances in phage display technology allow isolation of high affinity recombinant antibody fragments (Fabs) that may bind unique epitopes of biological threat agents. The versatility of the selection process lends itself to efficient screening methodologies and can increase the number of antigen binding clones that can be isolated. Pilot scale biomanufacturing can then be used for the economical production of these immunoglobulin reagents in bacterial fermentation systems, and expression vectors with hexahistidine tags can be used to simplify downstream purification. One such Fab reagent directed against botulinum neurotoxin A/B has been shown to be sensitive in a variety of assay formats including surface plasmon resonance (SPR), flow cytometry, enzyme linked immunosorbent assay (ELISA), and hand-held immunochromatographic assay. Recombinant antibodies can provide another source of high quality detection reagents in our arsenal to identify or detect pathogens in environmental samples.


Assuntos
Técnicas Biossensoriais , Fragmentos Fab das Imunoglobulinas/imunologia , Proteínas Recombinantes/imunologia , Animais , Afinidade de Anticorpos , Especificidade de Anticorpos , Toxinas Botulínicas/imunologia , Ensaio de Imunoadsorção Enzimática , Camundongos
9.
Artigo em Inglês | MEDLINE | ID: mdl-8861180

RESUMO

1. Phorbol 12-myristate-13-acetate (PMA), a protein kinase C activator, elevated cyclic AMP accumulation in EBV-transformed human B-lymphocytes, and potentiated isoproterenol-, prostaglandin- (PGE1), cholera toxin-, and forskolin-stimulated cyclic AMP accumulation. 2. The dopamine D1 receptor agonist, SKF38393 (10(-7) to 10(-5) MH, had no effect on cyclic AMP accumulation in transformed human B-lymphocytes. 3. The dopamine D2 receptor agonist, quinpirole (10(-7) to 10(-5) MH did not inhibit cyclic AMP accumulation even when cyclic AMP accumulation was maximized by the addition of PMA and forskolin. 4. These data suggest that dopamine D1- and D2-receptor coupling to a cyclic AMP generating system is not present at detectable levels in transformed human B-lymphocytes.


Assuntos
Alprostadil/farmacologia , Linfócitos B/efeitos dos fármacos , Toxina da Cólera/farmacologia , Colforsina/farmacologia , AMP Cíclico/metabolismo , Isoproterenol/farmacologia , Agonistas de Dopamina/farmacologia , Relação Dose-Resposta a Droga , Humanos , Ésteres de Forbol/farmacologia
12.
Exp Neurol ; 122(1): 48-56, 1993 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-8339789

RESUMO

To investigate the factors related to allogenic brain graft rejection, rat cortex from either LEW-RT1l or BN-RT1n rats was transplanted into brains of several isogenic and allogenic inbred strains: F344-RT1l, LEW-RT1l, BN-RT1n, AO-RT1u, PVG-RT1c, PVG-RT1u, and PVG-RT1l. Each donor and host combination was subsequently subdivided into two subgroups. One group was systematically sensitized twice with donor skin tissue and another group served as a sham-sensitization control. Four weeks after the second sensitization, host brains were examined histologically, and the percentage volume of each graft that showed increased cellularity was estimated. Expression of major histocompatibility complex (MHC) and T cell antigens was also studied immunocytochemically. Almost all grafts in sham-sensitized animals from all groups were accepted with minimal or no reaction. However, two strains (AO-RT1u and F344-RT1l) showed considerable cell infiltration and expression of MHC antigens even without sensitization. After sensitization, almost all allogenic strain combinations showed greater signs of rejection-related responses. The severity of the tissue reaction, however, varied considerably between groups. All grafts from BN-RT1n donors were rejected severely in all host strains. For LEW-RT1l donors, grafts survived well in some host strains (BN-RT1n, AO-RT1u, PVG-RT1c, and PVG-RT1u), even with MHC + non-MHC disparity. Curiously, F344-RT1l hosts rejected LEW-RT1l grafts even though the two strains have the same MHC loci, but different minor histocompatibility (mH) loci. For both donor strains, experimental autoimmune encephalomyelitis-susceptible hosts tended to show more vigorous rejection responses.(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Transplante de Tecido Encefálico , Rejeição de Enxerto , Imunização , Ratos Endogâmicos/fisiologia , Análise de Variância , Animais , Ratos , Ratos Endogâmicos/imunologia , Transplante Homólogo
13.
Am J Pathol ; 139(5): 1021-35, 1991 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-1951625

RESUMO

The major interacting components of the immune system, major histocompatibility complex (MHC) class I and class II proteins and T cells were analyzed in a model of anti-GBM (glomerular basement membrane) disease in the rabbit that progresses to develop cellular crescents and glomerular and interstitial fibrosis. Class I and II mRNA and protein were measured in isolated glomeruli and whole renal cortex using cDNA probes and monoclonal antibodies. The distribution of T cells and class I and II proteins was assessed by immunofluorescence. Normal glomeruli contained no T cells and were class II negative. By day 4, glomeruli contained MHC class I and II mRNA and protein and class II positive T cells. Although some animals had T cells in the periglomerular area, these cells were class II negative. By day 7 periglomerular T cells were largely class II positive (activated) and there was increased MHC class I and II mRNA and protein in whole renal cortex. Later T cells accumulated in the tubulo-interstitial compartment, which became diffusely positive for MHC classes I and II, but to a variable extent in different animals. Those with high class II mRNA expression also had detectable T cell antigen receptor mRNA by Northern analysis. The authors conclude 1) in this model there was a close association between mRNA abundance and protein expression for both MHC classes I and II in glomeruli and renal cortex as a whole; 2) in this model of glomerular injury there are three phases of activation. The first phase takes place in the glomerulus and is associated with accumulation of activated T cells and MHC class I and II protein in the glomerulus. Phase 2 is associated with the accumulation of periglomerular T cells and their becoming class II positive. There is subsequent dissemination (phase 3) of activated T cells and accumulation of class I and II mRNA and protein throughout the interstitial compartment. This spacial progression of glomerulocentric inflammation is likely associated with degree of injury and permanent loss of renal function.


Assuntos
Glomerulonefrite Membranosa/patologia , Antígenos de Histocompatibilidade Classe II/genética , Antígenos de Histocompatibilidade Classe I/análise , Antígenos de Histocompatibilidade Classe I/genética , RNA Mensageiro/análise , Linfócitos T/patologia , Animais , Anticorpos Monoclonais , Northern Blotting , Western Blotting , DNA/genética , Sondas de DNA , Modelos Animais de Doenças , Imunofluorescência , Expressão Gênica/genética , Glomerulonefrite Membranosa/genética , Glomerulonefrite Membranosa/imunologia , Antígenos de Histocompatibilidade Classe II/análise , Córtex Renal/química , Córtex Renal/patologia , Glomérulos Renais/química , Glomérulos Renais/patologia , Ativação Linfocitária/imunologia , Hibridização de Ácido Nucleico , RNA Mensageiro/genética , Coelhos , Receptores de Antígenos de Linfócitos T/análise , Receptores de Antígenos de Linfócitos T/genética , Linfócitos T/química
14.
Mol Pharmacol ; 50(5): 1309-15, 1996 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8913362

RESUMO

Dopamine produces a time- and dose-dependent increase in cell death in a clonal catecholaminergic cell line (CATH.a) derived from the central nervous system. Cell death also occurred after treatment with the catecholamines L-dihydroxyphenylalanine, norepinephrine, epinephrine, and isoproterenol, as well as the neurotoxic compound 6-hydroxydopamine. Cell death is not receptor mediated because selective noradrenergic and dopaminergic receptor agonists had no effect on CATH.a cell viability. Dopamine induces apoptotic cell death as indicated by DNA fragmentation measured by gel electrophoresis and by flow cytometric analysis. Apoptosis seems to be produced by dopamine autoxidation, because intracellular peroxides increase after dopamine treatment and cell death can be inhibited by catalase and N-acetylcysteine. N-acetylcysteine produced a dose-dependent decrease in dopamine-induced cell death; this correlated with a decrease in peroxide formation. In addition, antisense to the antioxidant protein bcl-2 increases the sensitivity of CATH.a cells to dopamine-induced cell death. These findings indicate that the oxidative products of dopamine cause neurotoxicity through apoptosis.


Assuntos
Apoptose/efeitos dos fármacos , Catecolaminas/fisiologia , Sistema Nervoso Central/citologia , Sistema Nervoso Central/efeitos dos fármacos , Dopamina/toxicidade , Animais , Neoplasias Encefálicas , Catecolaminas/farmacologia , Morte Celular/efeitos dos fármacos , Sistema Nervoso Central/metabolismo , Camundongos , Camundongos Transgênicos , Oligonucleotídeos Antissenso/farmacologia , Peróxidos/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Sensibilidade e Especificidade , Células Tumorais Cultivadas
15.
Proc Natl Acad Sci U S A ; 85(12): 4455-9, 1988 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-2454470

RESUMO

We report the successful infection of two rabbit T-cell lines and one rabbit macrophage line with human immunodeficiency virus 1 (HIV-1). One T-cell line was a herpesvirus ateles transformant, the other T-cell line was a human T-cell leukemia virus I transformant, and the macrophage line was a simian virus 40 transformant. After infection with a high-titered HIV-1 stock, the rabbit cultures exhibited properties that closely mimic those of HIV-1-infected human cells. Productive infection was evident in cultures 7-14 days after infection, as shown by an increase in reverse transcriptase activity, a concomitant increase in positive cells detected by indirect immunofluorescence using serum from a patient with acquired immunodeficiency syndrome, and a decrease in cell viability. RNA gel blot hybridization and protein immunoblot analyses of infected cells indicated that all predicted viral transcripts and proteins were synthesized during the course of the infection. Proof that cell-free culture supernatants of the infected rabbit cell lines contained infectious virus was given by successful passage onto a susceptible human T-cell line. The ability of HIV-1 to infect transformed rabbit cell lines in vitro suggests that, with appropriate manipulation, the rabbit may provide a model for infection with HIV-1.


Assuntos
Transformação Celular Viral , HIV/genética , Animais , Linhagem Celular Transformada , Imunofluorescência , Cinética , Macrófagos , RNA Viral/análise , DNA Polimerase Dirigida por RNA/metabolismo , Coelhos , Linfócitos T
16.
Res Immunol ; 140(5-6): 527-44, 1989.
Artigo em Inglês | MEDLINE | ID: mdl-2781136

RESUMO

Rabbits experimentally infected with HTLV-I and HIV-1 produced antibody to various viral proteins, and viral DNA could be detected by gene amplification using the polymerase chain reaction. HTLV-I genes were detected in cell lines derived from infected rabbits, and in some cases, both HIV-1 and HTLV-I DNA sequences were demonstrated in peripheral blood cells taken from rabbits one year after experimental infection. The polymerase chain reaction procedure was used to demonstrate the presence of HTLV-I gag, env and tax genes and HIV-1 gag and env genes. The amplified fragments were identified by size and by hybridization to specific probes. The ability of rabbits to support simultaneous infection with HTLV-I and HIV-1 will allow in vivo studies of the possible synergistic effects of these important human pathogens.


Assuntos
Síndrome da Imunodeficiência Adquirida/complicações , HIV-1 , Infecções por HTLV-I/complicações , Síndrome da Imunodeficiência Adquirida/imunologia , Síndrome da Imunodeficiência Adquirida/microbiologia , Animais , Linhagem Celular Transformada , DNA Viral/genética , DNA Viral/isolamento & purificação , Modelos Animais de Doenças , Amplificação de Genes , Genes Virais , Anticorpos Anti-HIV/biossíntese , HIV-1/genética , Anticorpos Anti-HTLV-I/biossíntese , Infecções por HTLV-I/imunologia , Infecções por HTLV-I/microbiologia , Vírus Linfotrópico T Tipo 1 Humano/genética , Coelhos
17.
Cephalalgia ; 18(4): 197-201, 1998 May.
Artigo em Inglês | MEDLINE | ID: mdl-9642494

RESUMO

We have conducted flow cytometric studies of two subsets of lymphocyte markers in groups of migraineurs during (n = 12; group B) and outside (n = 10; group C) of a migraine without aura attack (total n = 22; group A), including a group of patients tested in both of these phases (n = 5; group D), and compared these results with those obtained from a population of age-comparable, sex- and race-matched healthy volunteers (n = 12; group E). Comparison of the first set of lymphocytes (CD3+CD16 + 56+, CD3-CD16 + 56+, CD3-CD19+, CD3+CD19+, and CD3+HLA - DR+) between the patients in group A and the controls (group E) showed differences, reflecting greater group A percentages of CD3+CD16 + CD56+ and CD3-CD19+ lymphocytes. Furthermore, these differences reached statistical significance only for the CD3+CD16 + CD56+ lymphocytes, and then solely for the patients in group C (Scheffe's test, p < 0.05). Paired analysis of the above lymphocyte markers for subjects in group D failed to show significant differences between patients when they were having and not having a migraine attack, raising the possibility that results from a larger study could show meaningful increases in percentages of CD3+CD16 + CD56+ lymphocytes as one of the immune parameters useful for differentiating migraineurs from controls. Comparison of a second set of lymphocyte markers (CD19+CD5+, CD20+CD72-, CD20-CD72+, CD20+CD72+) among either the different groups of patients or between the patients and controls failed, however, to show statistically significant differences, emphasizing the apparent specificity of the findings described above for CD3+CD16 + CD56+ lymphocytes. Our results, albeit of a preliminary nature, suggest the occurrence of significant, differential changes in lymphocyte subset immunophenotyping between groups of pain-free migraineurs and patients during an acute migraine episode or controls. Corroboration of these findings may prove useful in clinical laboratory practice to identify changes in immunological parameters specifically associated with migraineurs, and help towards a better understanding of the etiology and pathophysiology of this condition.


Assuntos
Citometria de Fluxo , Subpopulações de Linfócitos/imunologia , Transtornos de Enxaqueca/imunologia , Doença Aguda , Adulto , Feminino , Humanos , Imunofenotipagem , Contagem de Linfócitos , Pessoa de Meia-Idade , Medição da Dor
18.
Am J Obstet Gynecol ; 158(3 Pt 1): 560-4, 1988 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-3348315

RESUMO

Deoxythymidine kinase is present in human cells in two forms: the cytosolar form (fetal thymidine kinase) and the mitochondrial form (adult thymidine kinase). The activity of thymidine kinase isozymes in plasma of preterm and term newborns has been previously reported to be related to gestational age, and fetal thymidine kinase activity was reported to be undetectable after the thirty-ninth week of gestation. Our purpose was to determine if similar changes in the activity levels of the thymidine kinase isozymes could be demonstrated in late third trimester maternal plasma. A cross-sectional sample of 35 patients was studied from week 34 of pregnancy to delivery. Significant differences between fetal and adult thymidine kinase activity patterns were observed. Adult thymidine kinase activity remained relatively constant throughout the evaluation interval, while fetal thymidine kinase activity decreased with advancing gestational age. Regression analysis of gestational age by neonatal examination (GAPED) and fetal thymidine kinase revealed a linear relationship (fetal thymidine kinase = 0.380 - 0.00745 GAPED; R2 = 0.2944; standard estimate of error = 0.0350; p less than 0.0008). Comparison of fetal thymidine kinase activity in term and postterm pregnancy plasma revealed a significant difference (p less than 0.0159). Our findings indicate that deoxythymidine kinase activity can be measured in maternal plasma, may relate to the fetoplacental growth rate, and may be useful in differentiating between term and postterm gestations.


Assuntos
Gravidez Prolongada/sangue , Timidina Quinase/sangue , Desenvolvimento Embrionário e Fetal , Feminino , Idade Gestacional , Humanos , Gravidez , Valores de Referência
19.
J Immunol ; 139(2): 587-92, 1987 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-3474276

RESUMO

Genomic clones corresponding to five distinct major histocompatibility complex class II alpha-chains have been described for the rabbit; four of these encode complete, potentially functional alpha-chains. Hybridization analysis and preliminary sequence analysis indicate that one of these clones is structurally related to HLA-DP alpha, one to -DR alpha, one to -DQ alpha and one to -DZ alpha. Probes specific for the four class II genes were used to screen RNA samples from normal rabbit tissues to determine which of these genes are transcribed and whether expression of any particular gene is tissue specific. All four of the genes are transcribed, but there are variations in the levels of expression, in tissue distribution, and in transcript size. The highest levels of RLA-DR alpha, -DQ alpha, and -DP alpha transcription were found in lymphoid tissues. Lower levels of transcription were also detectable in several nonlymphoid tissues. Transcripts observed were about 1.3 kb, a size expected for these class II alpha-chain genes based on experience with their human homologues. The RLA-DZ alpha probe corresponding to HLA-DZ alpha hybridized weakly with a band of 3.6 kb; its expression could be detected only in lymphoid tissues. The size of the DZ alpha transcript, its tissue distribution, and partial sequence data confirm its homology with the human gene DZ alpha. In blots of total cellular RNA, a probe for a recently described human beta-chain, DO beta, hybridized to a transcript of about 1.3 kb in lymphoid tissues. These data indicate that RNA transcripts corresponding to all HLA class II loci described to date can be detected in rabbit tissues.


Assuntos
Antígenos de Histocompatibilidade Classe II/genética , Complexo Principal de Histocompatibilidade , Coelhos/genética , Animais , Clonagem Molecular , Regulação da Expressão Gênica , Genes , Humanos , RNA Mensageiro/genética , Coelhos/imunologia , Distribuição Tecidual
20.
Exp Neurol ; 142(1): 179-94, 1996 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8912909

RESUMO

Olfactory neuroepithelial cells (ONC) grown from biopsies of human donors are a novel cell culture system that may facilitate studies into normal and disease-related human neurobiology. We further characterized the expression of cell surface markers and intermediate filaments, and responses to neurotrophic factors by ONC. ONC are positive for cell surface markers N-CAM, PSA-N-CAM, neutral endopeptidase, N-aminopeptidase, NGF low-affinity receptor homologue (CD40), and transferrin receptor by flow cytometry for the intermediate filament proteins peripherin, vimentin, and NF-H by immunocytochemistry. Responses to neurotrophic factors measured were process outgrowth, cytoskeletal protein expression, and protein phosphorylation. Process outgrowth was increased by interleukin-beta 164-171 (IL-1beta) or by the combination of IL-1beta, interleukin-6 (IL-6), nerve growth factor (NGF), and basic fibroblast growth factor (bFGF). This combination of IL-1beta, IL-6, NGF, and bFGF (16NF) increased expression of two cytoskeletal proteins, NF-H protein and microtubule-associated protein tau. Application of the individual neurotrophic factors IL-1beta, IL-6, NGF, and bFGF increased protein phosphorylation, while 16NF produced an immediate increase in tyrosine phosphorylation of several proteins (MW of 40-80, 120, 150, and 190 kDa). The 16NF combination appears to act through a tyrosine-kinase-mediated pathway to induce process extension and increase NF-H expression. The ONC culture has the potential to be further explored to examine the relationship among process outgrowth, protein phosphorylation, and synergy between neurotrophin and cytokine receptor systems.


Assuntos
Fator 2 de Crescimento de Fibroblastos/farmacologia , Interleucina-1/farmacologia , Interleucina-6/farmacologia , Fatores de Crescimento Neural/farmacologia , Condutos Olfatórios/citologia , Tirosina/metabolismo , Antígenos CD/análise , Antígenos de Diferenciação de Linfócitos B/análise , Biomarcadores , Antígenos CD13/análise , Antígenos CD40/análise , Diferenciação Celular/efeitos dos fármacos , Proteínas do Citoesqueleto/efeitos dos fármacos , Proteínas do Citoesqueleto/metabolismo , Epitélio/química , Epitélio/efeitos dos fármacos , Epitélio/enzimologia , Proteínas do Olho/análise , Citometria de Fluxo , Proteína GAP-43 , Proteínas de Ligação ao GTP/análise , Humanos , Imuno-Histoquímica , Proteínas de Filamentos Intermediários/análise , Glicoproteínas de Membrana/análise , Proteínas Associadas aos Microtúbulos/análise , Neprilisina/análise , Proteínas do Tecido Nervoso/análise , Moléculas de Adesão de Célula Nervosa/análise , Proteínas de Neurofilamentos/análise , Neuropeptídeos/análise , Mucosa Olfatória , Nervo Olfatório , Condutos Olfatórios/enzimologia , Neurônios Receptores Olfatórios , Periferinas , Fosforilação , Proteínas Tirosina Quinases/metabolismo , Receptores da Transferrina , Transdução de Sinais/fisiologia , Tirosina/efeitos dos fármacos , Vimentina/análise
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