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M Golightly

Publications and source records attributed to M Golightly.

8 recordsLinked to original sources
PubMed2000

Vitronectin is sequestered within human spermatozoa and liberated following the acrosome reaction.

Vitronectin plays a role in the regulation of complement and thrombin activities and in cell surface proteolysis. Vitronectin is also an intrinsic protein of human spermatozoa. Vitronectin message has been detected in whole testis poly-A mRNA and localized by in-situ reverse transcription-polymerase chain reaction to spermatocytes. The proportion of spermatozoa that express vitronectin increases following their capacitation. In this study, spermatozoa from a man of proven fertility were probed with an anti-vitronectin monoclonal antibody (VN7) before and after their permeabilization with 0.1% Triton X-100. Of fresh spermatozoa observed by confocal microscopy, 0-8% showed vitronectin staining. However, 75% of those observed displayed vitronectin following permeabilization. Serial confocal sections through the sperm head confirmed the internal localization of vitronectin. The acrosomal status of capacitated spermatozoa that expressed vitronectin was then determined. Dual colour microscopy with rhodamine-conjugated anti-vitronectin antibody and a fluorescein-conjugated antibody directed against CD46 (a complement regulatory protein expressed on the inner acrosomal membrane) revealed that only acrosome-reacted (CD46-positive) spermatozoa displayed vitronectin. Two populations of these spermatozoa were observed. Fifty-seven of 260 (22%) were CD46-positive/vitronectin-positive and 72 of 260 (28%) were CD46-positive/vitronectin-negative. No spermatozoa were CD46-negative/vitronectin-positive. These results confirm that vitronectin is released from a sequestered location within the spermatozoon following the acrosome reaction.

Acrosome Reaction↗
PubMed1999

Progesterone promotes the acrosome reaction in capacitated human spermatozoa as judged by flow cytometry and CD46 staining.

The acrosome reaction is a necessary prerequisite for spermatozoa to acquire fertilizing ability. Several different moieties appear to promote the acrosome reaction through different pathways, including solubilized zona pellucidae, recombinant zona protein ZP3, follicular fluid, calcium ionophores, and mannosylated bovine serum albumin (BSA). Although many investigators have presented evidence that progesterone also promotes the acrosome reaction through the mediation of a non-genomic cell membrane receptor, this concept has been challenged. Other workers have suggested that progesterone does not promote an acrosome reaction in human spermatozoa, as judged by the detection of CD46, a complement regulatory protein present on the inner acrosome membrane, through flow cytometric analysis of large numbers of spermatozoa. Prior investigations were criticized by the limited numbers of spermatozoa enumerated visually, the use of non-specific staining techniques, and the failure to eliminate dead spermatozoa during the scoring of the acrosome reaction. We have repeated these experiments, using both a supravital dye to eliminate dead spermatozoa from flow cytometric analysis, and anti-CD46 monoclonal antibody to score acrosome-reacted spermatozoa. Care was taken to validate the adequacy of capacitation conditions, which were proven by the ability of spermatozoa to acrosome react in response to mannosylated BSA and to penetrate zona-free hamster eggs. Confocal microscopy was used to confirm that CD46 immunostaining was limited to the acrosomal region of the spermatozoon head. Our results indicate that progesterone does promote an acrosome reaction within capacitated spermatozoa.

Acrosome Reaction↗
PubMed1994

Cell-mediated immunity in allopurinol-induced hypersensitivity.

Allopurinol may induce severe hypersensitivity characterized by hepatitis, interstitial nephritis, and skin rash. The mechanisms for this hypersensitivity syndrome are incompletely elucidated. Immunologic studies were performed on tissue and peripheral blood lymphocytes from a patient with allopurinol hypersensitivity. Immunohistochemistry was performed on sections of the liver biopsy utilizing monoclonal antibodies for T and B lymphocytes. Peripheral blood lymphocyte immunophenotyping by flow cytometry and peripheral blood lymphocyte stimulation studies with either allopurinol or oxypurinol measured as tritiated thymidine uptake were performed in the hypersensitive patient and compared to a group of six patients treated with allopurinol without hypersensitivity and eight normal control patients. Additional single- and dual-color immunophenotyping by flow cytometry of oxypurinol-stimulated lymphocytes was performed in the hypersensitive patient and compared to normal controls. The liver biopsy demonstrated predominantly a T lymphocyte infiltrate. The number of peripheral blood lymphocytes expressing activation antigens was significantly greater in the hypersensitive patient compared to that of both control groups. Lymphocytes from the hypersensitive patient were moderately stimulated by allopurinol and markedly stimulated by oxypurinol compared to both control groups. Oxypurinol-stimulated lymphocytes from the hypersensitive patient demonstrated enhanced expression of activation antigens compared to unstimulated lymphocytes from this patient and normal controls. These studies suggest that cell-mediated immunity directed toward allopurinol and more importantly to its oxypurinol metabolite is involved in the pathogenesis of allopurinol-induced hypersensitivity.

Acute Disease↗
PubMed1992

Lyme borreliosis in Bell's palsy. Long Island Neuroborreliosis Collaborative Study Group.

Lyme borreliosis (LB) causes a range of neurologic manifestations, the most common of which is facial nerve paralysis. To evaluate nervous system LB, we organized a neurologic collaborative study group in Suffolk County, NY, a region of high LB incidence. Between July and September 1989, LB serologies were performed on all patients with new-onset Bell's palsy. Seven of 32 had serologic evidence of LB at onset. One, initially seronegative, was highly seropositive 5 weeks later. In the five in whom we examined CSF, there was no evidence of intrathecal synthesis of specific antibody. In highly endemic areas, LB may be responsible for 1/4 of cases of Bell's palsy. Rarely, the palsy may occur prior to the development of a measurable antibody response, indicating a need for follow-up serologic testing.

Antibodies, Bacterial↗
PubMed1990

Immunologic reactivity against Borrelia burgdorferi in patients with motor neuron disease.

Of 19 unselected patients with the diagnosis of amyotrophic lateral sclerosis (ALS) living in Suffolk County, New York (an area of high Lyme disease prevalence), 9 had serologic evidence of exposure to Borrelia burgdorferi; 4 of 38 matched controls were seropositive. Eight of 9 seropositive patients were male (8 of 12 male patients vs 2 of 24 controls). Rates of seropositivity were lower among patients with ALS from nonendemic areas. All patients had typical ALS; none had typical Lyme disease. Cerebrospinal fluid was examined in 24 ALS patients--3 (all with severe bulbar involvement) appeared to have intrathecal synthesis of anti-B burgdorferi antibody. Following therapy with antibiotics, 3 patients with predominantly lower motor neuron abnormalities appeared to improve, 3 with severe bulbar dysfunction deteriorated rapidly, and all others appeared unaffected. There appears to be a statistically significant association between ALS and immunoreactivity to B burgdorferi, at least among men living in hyperendemic areas.

Adult↗
PubMed1988

Leukemia derived from intermediately differentiated lymphocytic lymphoma.

The availability of monoclonal antibodies has facilitated the immunophenotypic characterization of malignant lymphocytes from patients with lymphoma and leukemia. The chronic lymphocytic leukemias are diseases of both clinical and morphological diversity and the application of monoclonal antibodies can prove helpful in their classification. Enzyme cytochemistry, surface markers, mouse rosetting, and electron microscopy were used to determine the phenotype of cells from an atypical case of B-CLL. The use of monoclonals Leu-1, CALLA and BA-2 on bone marrow and peripheral blood provided the opportunity to diagnose this patient's disease as intermediately differentiated lymphoma. Leu-1 was found to be a useful alternative to mouse rosetting, a technique not easily performed in a routine setting. Ultrastructural studies helped to prove the prolymphocytic component of this patient's disease. It was concluded that phenotypic characterization of lymphoid cells using monoclonal antibodies directed against membrane antigens facilitated the assessment of this patient's disease.

Antigens, Surface↗
PubMed1980

Lymphocyte surface markers and cytotoxicity following cryopreservation.

The effects of cryopreservation (CP) on lymphocyte subpopulation distribution and functional activity in blastogenic and cytotoxicity assays were tested. Peripheral blood lymphocytes (PBL) from 12 healthy human donors were obtained by Ficoll-Hypaque separation. Half of each sample was tested fresh, while the other half was cryopreserved and then thawed and tested the same day. Each sample of CP-PBL was compared to fresh PBL from the same donor in simultaneous assays. Following CP there was a significant reduction in the percentage of E, EA gamma, and EA mu rosette-forming cells with a reciprocal increase in EAC rosette-forming cells. The blastogenic response to alloantigens was stable following CP while blastogenesis in unstimulated control cultures was significantly reduced. Mixed lymphocyte culture (MLC)-induced cell-mediated lympholysis (CML) was consistently and significantly diminished by CP. Cytotoxicity in 4 h chromium release NK (K562), ADCC, and LDCC assays was also significantly diminished by CP. In contrast, cytotoxicity was unaffected in an 18 h cytotoxicity assay against adherent cultured melanoma target cells.

Cell Survival↗