AW: AW: [Histonet] Helicobacter pylori immunocytochemistry - cytoplasmatic staining

[Gudrun Lang]

It is polymer. No endogenous biotin-problem.
The staining looks quiet specific. It is confined to cytoplasm. And there are clear negativ cells beside faintly stained "positiv" cells.
I'm reading about the envading features of helicobacter into host cells and about the "injection" of VacA and CagA into the host-cell via pili.
Therefore I think, the polyclonal antibody also detects those proteins. But this phenomenon should be a common thing?
Also cases are described, where the authors see an infra-nuclear area in gastric mucosa cells, that's metachromatic with Touluidinblue. (I'm not through the whole literature) But in their figures of IHC the intracellular stainings are stronger.
Perhaps the bugs change their shape and sit in the cells? Or the positiv cells mean something like: Helicobacter was here.

This night I repeat Hp-IHC with longer incubation and amplification to see, if the cellular staining can be enhanced, while the surrounding stays clear.

I work at a diploma-thesis with MALT-Lymphoma cases and Hp-detection. And now I came across this "funny" staining. I will also do a PCR on some of these cases. It will be interesting, if this staining is correlated with a positiv PCR.

Bye
Gudrun

-----Ursprüngliche Nachricht-----
Von: Richard Cartun [mailto:Rcartun <@t> harthosp.org] 
Gesendet: Mittwoch, 16. Jänner 2013 16:03
An: gu.lang <@t> gmx.at
Betreff: Re: AW: [Histonet] Helicobacter pylori immunocytochemistry - cytoplasmatic staining

Is that avidin-biotin based or polymer?

Richard

Richard W. Cartun, MS, PhD
Director, Histology & Immunopathology
Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-2204 Fax


>>> "Gudrun Lang" <gu.lang <@t> gmx.at> 1/16/2013 12:54 AM >>>
We work with ultraview-dab kit on ventana benchmark Ultra.

Gudrun

-----Ursprüngliche Nachricht-----
Von: Richard Cartun [mailto:Rcartun <@t> harthosp.org]
Gesendet: Dienstag, 15. Jänner 2013 23:44
An: gu.lang <@t> gmx.at
Betreff: Re: [Histonet] Helicobacter pylori immunocytochemistry - cytoplasmatic staining

Which Ventana detection system are you using?

Richard

Richard W. Cartun, MS, PhD
Director, Histology & Immunopathology
Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-2204 Fax


>>> "Gudrun Lang" <gu.lang <@t> gmx.at> 1/15/2013 3:56 PM >>>
Hi!
I need some help with the interpretation of Hp-IHC. I stained several cases of FFPE gastric biopsies with polyclonal anti Hp from Cell Marque (1:200, ventana benchmark).
Some cases showed the nice bacteria and had usually a clean overall background. But some cases showed no bacteria but slight cellular staining of epithelial cells. 
It was confined on the cells, and usually not all cells in the biopsy had this staining.

>From literature I learned, that Hp brings its toxine and other
proteins
(CagA) into the hostcell. Is it possible, that the polyclonal antibody detects this stuff, although hardly an intact bacterium can be seen?
Anybody with similar findings?

Thanks in advance
Gudrun Lang


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