scholarly journals Influence of two different fixatives on the identification of plasma cells in human rectal mucosa.

1980 ◽  
Vol 28 (9) ◽  
pp. 1018-1020 ◽  
Author(s):  
M F Heyworth
Keyword(s):  
Plasma Cells ◽  
Rectal Mucosa ◽  
Rectal Biopsy ◽  
Immunoperoxidase Technique ◽  
Biopsy Specimens ◽  
Indirect Immunoperoxidase

Plasma cells in sections of bisected human rectal biopsy specimens, fixed in two alternative fixatives, were enumerated after staining by an indirect immunoperoxidase procedure intended to demonstrate immunoglobulin-containing cells. The counts of immunoperoxidase-positive plasma cells were significantly higher after fixation in formol sublimate than after fixation in formol saline. Formol sublimate appears to be a more reliable fixative than formol saline for specimens of rectal mucosa in which quantitation of plasma cells, stained for intracellular immunoglobulin by an immunoperoxidase technique, is intended.

scholarly journals 3160 The Effects of Aging on the Rectal Mucosal CD4+ T cell Compartment and its Implications for HIV Transmission

2019 ◽  
Vol 3 (s1) ◽  
pp. 115-116
Author(s):  
Cassie Grimsley Ackerley ◽  
Praveen Kumar Amancha ◽  
Phillip M. Murray ◽  
Jasper Barnes ◽  
Colleen F. Kelley
Keyword(s):  
Target Cell ◽  
Hiv Transmission ◽  
Rectal Mucosa ◽  
Rectal Biopsy ◽  
Study Groups ◽  
Cd4 T Cell ◽  
Hiv Susceptibility ◽  
Biopsy Specimens ◽  
Effects Of Aging ◽  
Hiv Negative

OBJECTIVES/SPECIFIC AIMS: In the first aim, we will evaluate the proportion of highly HIV-susceptible memory CD4+ T cells present in the rectal mucosa, based on the proliferation status and expression of the HIV susceptibility markers, CCR5 and α4β7, between HIV-negative adolescent MSM and adult MSM engaging in RAI. The second aim will assess differences between the two study groups in the ratio of Th17 cells (CD4+ IL17+) to Treg cells (CD4+ FoxP3+ CD25+) in the rectal mucosa as a determinant of mucosal inflammation. Finally, in the third aim, we will utilize ex vivo rectal biopsy explant challenge experiments to examine whether HIV target cell availability and the Th17/Treg ratio influence rectal mucosal HIV susceptibility. METHODS/STUDY POPULATION: Rectal biopsy specimens are being collected from healthy, HIV-negative men that comprise the two study groups: 40 adolescent MSM 18-21 years of age who have engaged in RAI at least once previously in their lifetime and 40 adult MSM ≥35 years of age who have engaged in RAI for the previous 5 consecutive years with a minimum of 12 episodes annually. To identify CD4+ subsets of interest for aims 1 and 2, rectal mucosal mononuclear cells are isolated and phenotyped with CD45, CD3, CD4, CD45RA, CCR7, CD69, CCR5, α4β7, Ki67, FOXP3, and CD25 antibodies. To identify the Th17 cell subtype, the cells are stimulated with PMA/Ionamycin and stained with an antibody specific to IL-17A. Using cross-sectional analyses, we will compare the frequencies of mucosal CD4+ T cells that express certain phenotypic characteristics and evaluate differences in the Th17/Treg ratio between adolescent and adult MSM. For aim 3, rectal biopsy specimens are inoculated with HIV virus and the culture supernatant is assayed for p24 concentration on days 3, 7, 10 14, and 18. Longitudinal analyses will be performed to detect differences in p24 concentration at each time point and assess associations with mucosal target cell availability and with the Th17/Treg ratio. RESULTS/ANTICIPATED RESULTS: We hypothesize that younger age will be associated with enhanced memory CD4+ T cell proliferation and increased expression of HIV susceptibility markers (CCR5 and/or α4β7). In addition, we expect that the rectal mucosa of adolescent MSM will demonstrate a higher Th17/Treg ratio as compared to adult MSM, which could facilitate HIV transmission. It is also anticipated that rectal mucosal immune phenotypes characterized by increased HIV target cell availability and high Th17/Treg ratios will be associated with enhanced mucosal HIV susceptibility in the explant challenge model. DISCUSSION/SIGNIFICANCE OF IMPACT: There is a paucity of information regarding the mechanisms of rectal HIV transmission, and no studies to date investigate the immunologic effects of aging on transmission in the rectal mucosa. The results from this study will provide important information regarding age-related differences in the immune cell composition of the rectal mucosa as a critical step in better understanding immunologic factors that influence rectal HIV transmission.

scholarly journals Gliadin activates mucosal cell mediated immunity in cultured rectal mucosa from coeliac patients and a subset of their siblings

Gut ◽  
1998 ◽  
Vol 43 (4) ◽  
pp. 484-489 ◽  
Author(s):  
R Troncone ◽  
G Mazzarella ◽  
N Leone ◽  
M Mayer ◽  
M De Vincenzi ◽  
...  
Keyword(s):  
Rectal Mucosa ◽  
Rectal Biopsy ◽  
Cell Mediated Immunity ◽  
Mucosal Cell ◽  
In Vitro System ◽  
Biopsy Specimens ◽  
Activate Cell ◽  
Cell Adhesion Molecule 1

Background—CD3 and γδ cells in the rectal mucosa increase after local instillation of gluten in children with coeliac disease and in half of their siblings.Aim—To establish an in vitro system for assessing immunological changes induced by gluten in the rectum.Patients and Methods—Rectal biopsy specimens obtained from 13 treated coeliac children, nine of their siblings, and nine controls were cultured in vitro with a peptic-tryptic digest of gliadin or ovalbumin. CD3 and CD25 cells were counted, and the expression of adhesion molecules evaluated.Results—In the lamina propria of coeliac biopsy samples cultured with gliadin, but not in those from controls, the expression of vascular cell adhesion molecule 1 (VCAM-1) was enhanced, and the number of CD25 cells was significantly higher than in those cultured in medium alone; the density of intraepithelial CD3 cells was also significantly higher. No differences were noted in coeliac biopsy specimens cultured with ovalbumin. A discriminant analysis allowed correct classification of all controls and all coeliacs but one, but three of nine siblings were allocated to the coeliac group.Conclusions—Our data confirm that gliadin is able to activate cell mediated immunity in the rectal mucosa in coeliac patients and in a subset of their first degree relatives.

scholarly journals A novel tetrazolium method for peroxidase histochemistry and immunohistochemistry.

1991 ◽  
Vol 39 (4) ◽  
pp. 541-544 ◽  
Author(s):  
G I Murray ◽  
C O Foster ◽  
S W Ewen
Keyword(s):  
Peroxidase Activity ◽  
Histochemical Demonstration ◽  
New Method ◽  
Tetrazolium Salt ◽  
Immunoperoxidase Technique ◽  
Red Cell ◽  
Final Reaction Product ◽  
Biopsy Specimens ◽  
Oxidation Of Phenol ◽  
Indirect Immunoperoxidase

We developed a new method for the histochemical demonstration of peroxidase. This method, which has a novel reaction mechanism, is based on the oxidation of phenol by peroxidase and coupling of this reaction to the reduction of a tetrazolium salt, with the deposition of an insoluble formazan at sites of enzyme activity. This new method was compared with an established diaminobenzidine (DAB) technique for peroxidase histochemistry and immunohistochemistry. Although both methods identified peroxidase activity in myeloid cells of bone marrow biopsy specimens, there was no interference from red cell pseudoperoxidase activity with the phenol-tetrazolium method, in contrast to the diaminobenzidine method. The detection of cytokeratin using an indirect immunoperoxidase technique was compared with both methods for demonstrating peroxidase activity. The phenol-tetrazolium method gave results similar to that obtained with DAB and appeared to be at least as sensitive as DAB in detecting low amounts of antigen. In addition, the production of a formazan as the final reaction product means that the phenol-tetrazolium method is ideally suited for quantitative peroxidase histochemistry. Therefore, the phenol-tetrazolium method represents a useful alternative method to DAB and for certain applications offers significant advantages over DAB.

scholarly journals Varicella-zoster plaque assay and plaque reduction neutralization test by the immunoperoxidase technique

1976 ◽  
Vol 4 (5) ◽  
pp. 437-442
Author(s):  
G Gerna ◽  
R W Chambers
Keyword(s):  
Neutralization Test ◽  
Plaque Assay ◽  
Neutralizing Antibody ◽  
Immunoperoxidase Technique ◽  
Serum Samples ◽  
Plaque Reduction Neutralization Test ◽  
Varicella Zoster ◽  
Convalescent Phase ◽  
Antibody Titers ◽  
Indirect Immunoperoxidase

A new plaque assay for the quantitation of varicella-zoster virus and a plaque reduction neutralization test for the determination of neutralizing antibody titer have been developed using the indirect immunoperoxidase technique. As compared with the classical plaque assay using a solid overlay, the test gives earlier results since plaque counting can be performed on day 3 after the inoculation of cell cultures. In six patients with zoster infection, neutralizing antibody titers ranged from 1:20 to 1:40 before the onset of infection and reached high levels (1:320 to 1:5,120) during the convalescent phase of the disease. Complement-fixing (CF) titers were all negative (less than 1:8) in prezoster serum samples from the same patients and ranged from 1:128 to 1:2,048 in the convalescent-phase sera. In the two cases in which late serum samples were available, neutralizing antibody titers matched the preillness levels, whereas CF titers dropped to undetectable levels. Neither neutralizing nor CF antibody was detected in two sera from individuals with no history of varicella-zoster infection. No differences in virus titers or neutralizing antibody titers were observed between the immunoperoxidase and the classical plaque assays. The appropriate characterization of reagent specificity is required before routine application of the test.

Total Colonic Aganglionosis: Case Report, Practical Diagnostic Approach and Pitfalls

2010 ◽  
Vol 134 (10) ◽  
pp. 1467-1473 ◽  
Author(s):  
Raja Rabah
Keyword(s):  
Transition Zone ◽  
Ganglion Cells ◽  
Hirschsprung Disease ◽  
Rectal Biopsy ◽  
Nerve Fibers ◽  
Submucosal Plexus ◽  
Total Colonic Aganglionosis ◽  
Biopsy Specimens ◽  
Suction Rectal Biopsy ◽  
Acetylcholinesterase Staining

Abstract Hirschsprung disease remains a challenging diagnosis for many pathologists. The disease is characterized by a lack of ganglion cells in the myenteric and submucosal plexus, associated with increased numbers of acetylcholinesterase-positive nerve fibers. Hypertrophic nerve fibers are present in most but not all patients. Total colonic aganglionosis (TCA) is an uncommon form of Hirschsprung disease with clinical, histologic, and genetic differences and is even more difficult to diagnose and manage. This case illustrates some of the difficulties frequently faced by the pathologists dealing with total colonic aganglionosis. Suction rectal biopsy specimens often lack significant nerve hypertrophy and positive acetylcholinesterase staining, which aid in the diagnosis. Pathologists have to depend mainly on the lack of ganglion cells in adequate submucosa to establish the diagnosis. Transition zone is often long in total colonic aganglionosis and interpretation of frozen sections can be difficult. The presence of several uniformly distributed clusters of mature ganglion cells and lack of nerve hypertrophy are required to avoid connections at the transition zone.

Megacystis-Microcolon-Intestinal Hypoperistalsis Syndrome

PEDIATRICS ◽  
1979 ◽  
Vol 63 (5) ◽  
pp. 805-808
Author(s):  
Thomas E. Wiswell ◽  
James S. Rawlings ◽  
James L. Wilson ◽  
Gary Pettett
Keyword(s):  
Neonatal Period ◽  
Ganglion Cells ◽  
Plain Film ◽  
Rectal Biopsy ◽  
Exploratory Laparotomy ◽  
Nerve Fibers ◽  
Bladder Obstruction ◽  
Distended Abdomen ◽  
Biopsy Specimens ◽  
Hypoperistalsis Syndrome

The syndrome of megacystis-microcolon-intestinal hypoperistalsis has been reported in a total of seven female infants. Massive abdominal distention secondary to a distended urinary Fig 1. Plain film of markedly distended abdomen with no air beyond stomach. Stomach is displaced upward toward left. der was the major presenting characteristic. Iutestinal hypoperistalsis, apparent in the early neonatal period, persists without improvement. Exploratory laparotomy reveals malrotation and malfixation of a small microcolon. No anatomic cause of intestinal or bladder obstruction can be found. Intestinal and/or rectal biopsy specimens contain abundant ganglion cells and nerve fibers. The outcome has been uniformly fatal, with survival in the reported cases ranging from two days to 34 months.

scholarly journals Evidence for significant hematopoiesis in the human thymus

Blood ◽  
1976 ◽  
Vol 47 (2) ◽  
pp. 305-313 ◽  
Author(s):  
CR Taylor ◽  
JM Skinner
Keyword(s):  
Plasma Cells ◽  
Positive Staining ◽  
Immunoperoxidase Method ◽  
Thymic Tissue ◽  
Fetal Thymus ◽  
Neonatal Deaths ◽  
Hemoglobin A ◽  
Staining Techniques ◽  
Immunohistochemical Methods ◽  
Indirect Immunoperoxidase

Abstract Earlier studies on fetal thymus suggested that certain of the large pyroninophilic cells found there might have a hemopoietic role, and it was decided to determine the nature of these cells using histochemical and immunohistochemical methods. Thymic tissue from aborted fetuses, stillbirths, and neonatal deaths was examined histochemically using methods for the detection of chloroacetate esterase, peroxidase, and pseudoperoxidase, and by staining techniques for mast cells and eosinophils. Tissue was also examined using the indirect immunoperoxidase method for the presence of hemoglobin A (HbA) and F (HbF), for lysozyme (muramidase) and immunoglobins alpha, mu, gamma, kappa, lambda. Positive staining to some degree was seen in cells in the connective tissue stroma using all methods, and the cells stained corresponded to one or another of the types of pyroninophilic cells present. The finding of large cells with positive chloroacetate esterase and antilysozyme indicates the presence of granulopoiesis. Similarly, the presence of large nucleated cells with pseudoperoxidase and anti-hemoglobin (A and F) staining indicates the presence of erythropoiesis. Plasma cells were present in small numbers.

scholarly journals Depletion of neuroendocrine cells in rectal biopsy specimens from HIV positive patients.

1992 ◽  
Vol 45 (6) ◽  
pp. 524-527 ◽  
Author(s):  
J B McCullough ◽  
P A Batman ◽  
A R Miller ◽  
P M Sedgwick ◽  
G E Griffin
Keyword(s):  
Rectal Biopsy ◽  
Neuroendocrine Cells ◽  
Hiv Positive ◽  
Biopsy Specimens
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