Oncology lab question - stains & PCR when and why

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Gator & H (GA)

Gator & H (GA)

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F [H's brother] got a needle aspirate from his spleen or somewhere around his spleen. The cells were "abnormal" which doesn't say much as I understand since spleen cells can typically be abnormal. Only after we asked them if stains or PCR testing on the sample would be any help they said, "yea we can send the sample out for stains then we would KNOW if it is cancer or not." Well duh!!! Uggg. They did not seem to really know/respond to the PCR testing thing.

Question:
When are stains and or PCR testing on a needle aspirated sample warranted? Should stains be done before PCR testing is done? Or should one just have PCR testing done on the sample and not even worry about the stains? I guess what I'm asking is at an individual level, at what point are stains and PCR testing warranted on an oncology sample?

As H's other brother S was killed by a misdiagnosed positive needle aspirate [at this same institution F is at currently] my faith in needle aspirates is already ultra-low. :-x I posted in the Yahoo lymphoma group too. But I've seen a few people around here that really knew their oncology stuff. :smile:

Any other input is welcome. Thank you for your replies/input/opinions.
 
To interpret aspirates some of the material is placed on a slide and the slide stained. I am not aware of any real meaningful analysis with staining. However, in some instances special staining is done. Regarding PCR testing, I Googled for "PCR test for feline cancer" and really only get a PCR test for Leukemia.

Very frequently aspirates are inconclusive because the small sample just does not contain the "right" cells. Typically the slides of aspirates (and biopsies) are interpreted/reviewed by pathologists because they have the experience of looking at slides of aspirates and biopsies. Who reviewed/interpreted the aspirate slides?
 
Larry,
Supposedly "3 people" looked at the sample - hopefully a pathologist was at least one of these 3. We're finding out currently if in fact a pathologist looked at the sample - I think that was your question. If not then I'll post again. I'm also in the process of trying to get instutution 1 to talk to institution 2 to see if institution 2's lab can help - uggg.

You might do better if you search "clonality test" - I'm assuming clonality testing = PCR? The way I know about PCR is from the other institution I took H to north of me. I was pretty sure he had PCR testing done on his lymphoma samples. From H's pathology report:
The neoplastic round cell population expanding the jejunal lamina propria was immunoreactive for CD3, which is consistent with T-cell population. Clonality test confirmed the diagnosis of T-cell lymphoma with clonal TCRG rearrangement.
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Do you know how immunoreactivity is determined?
 
Hi Gator

I think you're thinking of flow cytometry. This can help determine the B-cell, T-cell population in a bunch of abnormal lymphocytes. Can you post the text from the cytology report? Stains can help if they are suspecting certain things like parasitic infiltration, but flow cytometry is the way to go if they are looking for lymphoma type. FWIW, lymphoma is fairly easy to see on cytology. Perhaps if you post the report you got we can see what they are thinking of.

Also, I'm not sure what you mean by abnormal cells being normal for the spleen? The spleen can look funny on ultrasound for benign reasons, not sure if that is what you mean, but a cytology should distinguish between benign and malignant changes.
 
PCR = Polymerase chain reaction which is used to amplify strands of DNA to identify infectious agents .
i am guessing the PCR in reference to leukemia( larry's post) was for FLV.

Winnie had immunohistochemical testing on her biopsy. have been up all night.
but i can dig it up later if you like and see what jogs my brain.

chriscleo had cleo's spleen aspirated and tested and I *think* finally removed.
You might check with her -- she went through a lot w/ her vets on spleen issue w/ cleo's lymphoma.

sorry your kitty is sick.
 
OK Wanted to give you guys the update.

The initial pathology for F said:
Interpretation:
Lymphoid proliferation; Mild neutrophilic inflammation; Mild mastocytosis

Comment:
The lymphoid proliferation raises a serious concern for lymphoma ...
Click to expand...
There was a little more after that, including the suggestion that the sample be analyzed via "histopathology." Imagine that!

Sample had immunohistochemistry done on it [since it is from needle aspirate - otherwise immunohistopathology for full biopsy I guess]. Sample had anti-CD3e T and anti-CD79a B cells - "no strong evidence for neoplasia."

I had a heck of a time running down where to have this accomplished. The sample has not yet been sent there yet and the story is not over. I'm still waiting to find out of PCR/Clonality would be appropriate for in this case. I would think it to be quite astonishing if this PCR/Clonality testing could take an ambiguous sample from fine needle aspirate and add a layer of certainty to it. However, even if no evidence for lymphoma were found this is just from a needle aspirate which is limited in its sampling ability. But if something came back as an absolute for lymphoma then that would be beneficial and we would not have to do laparotomy/explatory to find out if he had lymphoma.
 
Hi Gator

So for the flow cytometry staining you'd be looking for either a certain pattern or (more commonly) a strong over-representation of either cell population. YOu've only posted a snippet, but with a low count mixed population it doesn't sound too exciting.

For reference, U of Illinois and UC Davis (Dr. Valli) are both considered great centers for diagnosis of lymphocyte-related disease. If you do another aspirate you can submit directly to them. Colorado State (CSU) also does the PARR (clonality) testing but I haven't heard about them one way or the other. The best veterinary cancer pathologist in the country, and probably (no exaggeration) the world, is Dr. Barb Powers at CSU, but she only works with tissue samples, no aspirates -- she is the last stop for any head-scratching cases. Hopefully no one will ever need this info, but I'm putting it out there.

To answer your original question, Gator, the extensive additional testing is warranted whenever there is serious suspicion of cancer. This means either symptoms (weight loss, LN enlargement, blood count disorders, etc.) or just plain no other explanation for an abnormality, esp. in an older cat. I don't know if your cat has any symptoms, but if there was no other explanation offered for the appearance of the spleen (was that why you did the aspirate? or size, or something?), it'd be worth looking at again ~3 weeks later to look for changes and, if warranted, get another aspirate. If he does unfortunately have lymphoma seeded in the spleen (which sounds unlikely given your results so far), you should see advancement of the disease. (Some vets would say wait a month, or 2-3 months, but I am not one to sit on my hands for so long!)

Hope this helps....
 
Hi Jess, yes it is a wonderful help.

And yes, F is in sorry shape. As F has not been [and is not currently] in my care, things have gotten a little too far [and maybe un-recoverably] out of control. And unfortunately everything I have to report is 3rd party. Very skinny, inappetence, CRF, Spec fPL high but not crazy high [at 10 - 0.1-3.5 is normal], but not I don't think I read about enlarged LNs in his reports. Past endoscopies have reported IBD but no lymphoma. Via PCR he does not have Helicobacter [or any strain]. MRI reports no significant neurological abnormalities. He's finishing antibiotics for potential infection around pancreas area, getting fluids and appetite stims. No improvement has been noted with attempted pain relief via Torbutrol - but he does not look/act in pain and is not meatloafing/hiding/etc.

Thank you for your wonderful compendium of other resources for diagnosis of lymphocyte-related disease. Have you committed this info to something like peddiabetes wiki? It would be a little off topic but... PDW does not seem to have a lymphoma section?? Was also thinking "PCR" probably deserves it's own explanation too in PDW. I tried to add a little under the IBD portion but this technology has so many incredible uses that it seems many vets are not even aware of and it takes owners to advocate for it.
 
Hi Gator,
this whole discussion is over my head but just wanted to send you hugs. hope you find the results your looking for. and that your doing ok.
Lori
 
Thanks Lori. Your thoughts are very appreciated. As this is not "my" cat I'm fairly emotionally OK with it. The caregiver is a parent though who refuses [and has reused now for a long time] to get on the [..er or should I say *my*] bandwagon in many regards about his care which is my largest emotional issue going on. *sigh* The other option is to bring him out here and me care for him which I'm not sure I'm really ready for at this point. This cat has lived a great life indoors/outdoors and sticking him in my little place here I don't think is much of a solution [for him or me]. He's the only one of the five brothers to live this long. No good choices so far with this.
 
don't know how old kitty is or how far away he lives but the move could be a trauma. is he in pain? is he loved? is his life pretty decent?
not all kitties get the 5 star treatment gator, it's a hard cold fact. all of our kitties are extraordinarily lucky.
check out my latest heart attack with tom on pzi tonight. i can''t count how many trauma's my boy has given me since gracing me with being his entire staff 6 years ago. the trauma's only seem to make us closer tho'.
lori
 
Thought I would give an update to this topic. Today we got the definitive news.

Short story first:
Though PCR/clonality testing of older samples from 2009 endoscopy, F has small [assumed T cell] GI lymphoma.

Now the long story:
At my instance, PCR clonality testing was done on a needle aspirate sample from his spleen. The institution where the original cytology was done did not even know PCR could be done for lymphoma. :roll: Results on that spleen sample were still somewhat inconclusive - but did indicate suspected LGL on visual inspection by the PCR lab/institution [not the original institution]. Somehow the idea was hatched to get old full tissue samples from Antech of F's old endoscopies from 2008 and 2009. Antech at first refused to send the full tissue samples to the PCR lab. It took us nearly two weeks of moving mountains with Antech [raising Cain and showing fangs] to get F's paraffin block samples sent to the PCR lab. The amount of time they wasted with the originating vet and our time they wasted was shameful. Finally, clonality tests were run on the 2009 GI samples. Below are the report "comments":
Molecular clonality analysis of TCRG (T cell) revealed a clonal rearrangement. In conjunction with the histopathological changes, the lesion is interpreted to be neoplastic in nature, most likely of T cell origin. The intestinal sample was run alongside the splenic sample evaluated in September. Peak sizes were different suggesting the presence of two separate neoplastic processes. The most likely scenario is that there is that there may be a more than one T cell lymphoma in the intestine - the other at a site not biopsied - this has been seen several times in our case series of intestinal lymphomas. The less likely option is that the splenic lesion is a primary lymphoma - this is far less common in our experience, but still a valid option.
Click to expand...
Antech pathology did not diagnose lymphoma in 2009. My understanding is that some pathologists may not currently diagnose lymphoma confined to the mucosal layer of the intestine - that they look for transmural progression - and that this modal of thinking may be changing.

I think the lesson here for everyone is that PCR is a powerful new tool for the diagnostic of lymphoma. Further, there are only currently isolated places [like Michigan State, perhaps NC State, UC Davis and perhaps others] where this is done and usually only by referral. IMHO, lymphoma suspect pathology samples should be sent to a lab with this PCR capability, perhaps even directly forgoing the local or 'normal' labs. Positive or negative mis-diagnosis of this disease has played a significant or life and death roll now in two cats from this litter. I'm very disappointed that I [the non-vet/expert] had to be the 'expert' in the case to get F's tissue samples tested via PCR. My insistence on PCR has hopefully helped avoid further pain and distress from the recommended exploratory surgery to an already very frail [1.5 body condition] and sick kitty. In this case, the additional cost of the PCR was perhaps 1/50th the expected cost of exploratory surgery and it is available to anyone who asks for it. In the end, I consider this a 'happy' ending - at least we know what we are dealing with and have potentially avoided further interventions for the sake of diagnosing lymphoma.

Secondary lesson:
The owner of tissues samples is the owner of the animal [unless ownership has been relinquished to another party for some strange reason]. Pet owners have the right to have done as they desire with the samples wherever they reside. Do not let some lab buffalo you into doing as they please with your samples! Do not take "no" for an answer.

I'll be more than happy to help anyone resource getting potential lymphoma samples tested for PCR.
 
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