r/Immunology

Hi fellow immunologist. I am having some viability issues with murine CD8 culture that I am looking for help with. Here are my general steps.

1. Negative selection of mouse splenocytes using the miltenyi CD8 isolation kit. Acquiring 4-12 million CD8s per spleen with approx 100% viability.

2. Either

(a) seed cells in wells precoated CD3/28 or another TCR engager (standard 2 hour coating protocol, using 20-50nM concentration) or

(b) treat CD8s with soluble TCR engagers (including an anti-hamster antibody for anti-CD3/28 to support crosslinking). In each condition, I have unstimulated and IL2 controls.

3. 24h later, analyze cells.

My issue is that my viability is generally poor upon viability analysis after 24h culture, between 20-50% following this 24 hour incubation, across all treatments (with and without TCR engager, with IL2 or not). I have altered seeding density (1-2.5 million per mL), used flat-bottomed or round-bottom culture (the latter only with soluble engagers), and currently use the following media formulation: RPMI with L-glut, 10% FBS, 1% pen-strep, 25uM 2-ME.

Will any of you fine folk share your thoughts and experiences?

Hi everyone! I have a presentation due TOMORROW that I just remembered about gamma delta T cells and I don’t have time to read all the articles i saved!! Help

I’m confused about how it activates and effector functions. Does it come pre activated from the thimus and then it activates due to stress, damaged cells and stuff?

Is this the part where it “decided” which cytokine is going to express?

Also effector functions?? Help lol

Do you think there is something i should not forget to talk about? Just so i can check my slides

Thank you

Hello everyone. I’m a medical student and I have finished Abbas’ Basic Immunology and Abbas’ Cellular and Molecular Immunology. To go further, I would like to read another Immunology book. What book do you recommend for my level?

Hi,
Does anyone have experience to freeze cultured T cells in 10% DMSO + 90% Human serum?
We are usually using a freezing medium with FBS and RPMI, but have to go away from FBS and collaborator suggested 90% HS.
Experience with long-term storage (up to several years) of T cells in this media?

please share with source?

In 2022 I did a standard std postal test where I tested negative for HIV after submitting a finger prick sample of blood

In July 2023 I did another kit and the finger prick came back as reactive , I went to the clinic where they took blood directly from the vein and did the rapid test there which again was reactive. They sent the vein blood to the lab for confirmation. The first antibody was reactive again but confirmation was all negative.

September 2023 I had blood test direct from vein and the same thing - screening was reactive followed by negative confirmation. This was repeated in October 2023 with the same results: reactive and then negative confirmation.

Then April 2026 direct from the vein is reactive followed by negative confirmation tests. This was repeated start of April and the end of April.

So in total there has been 7 reactive screening tests followed by 6 negative confirmations. This is across 3 years, 4 different labs.

I did not do another test between 2023 and 2026 as I was completely traumatised by the experience (due to the first ever test being an unknown false positive and receiving no guidance) and no healthcare can advise me on what the cause is.

I went to my GP recently where they screened for a few different things and full blood count and everything seems normal apart from low iron and low retocytes? They screened for RA as it is a common cause and in my family but this was not indicated from the tests

Whenever I ask gp and sexual health about this they don’t give me info on the next steps regarding how I can find out what is causing this false reaction.

My main concern is that before 2023 i had various standard hiv screenings and I did not test reactive. Clearly it isn’t a lab issue or linked to a flu as this is a repeated result for 3 years now.

What could this be and where can I find more answers, my GP said that they don’t have a crystal ball to know what’s causing it. Obviously my main concern is that I have something underlying / asymptomatic auto immune disease ? I 100% don’t have HIV, at extremely low risk and have not had any symptoms so I’m accepting with the fact it is a genuine false reaction and I don’t have HIV.

I feel like I need to speak to someone / advice form a lab tech who preforms these tests but I have no idea where to find help with this or where I can ask my GP to refer me to?

Streptococus neumoniae has more than 100 variants. I want to know the effects of getting vaccinated against all of them the same day. ( if you know about other bacterias or viruses is useful too ).

Im interesting on the effects of ussing a large number of vaccines at the same time. Does the inmune system manage to generate an adequate response to all of them? Do we lose effectiveness? Any reason not to do it if an individual can afford it?

I assume the prevalence of side effects increases but i cant find nothing about a limit on the number of vaccines per day (or short period of time).

I am looking to get Janeway's and Kuby Immunology textbooks. I am looking to get physical copies - I have also realised I learn better that way and with the amount of screen time i consume I'd much prefer a physical copy. I am a PhD student on a budget, and dont mind thrifted or used books.

EDIT: Based in Houston, Texas.

Does anyone know where I could get some work experience/placement or an internship for immunology and pharmacology related companies. As I have searched everywhere and they all say the application window has closed, I have been on the lookout for a while and it always takes me to the application window has been closed anytime I try.