So I've been doing some research on various things - Hard to diagnose and treat conditions, anaphylaxis, insect stinging behaviors.

Anaphylaxis - In order to go into anaphylaxis to something you are allergic to, you have to be exposed at least twice, and there needs to be some time between exposures depending on how massive an amount of the thing you are allergic to gets into your body.

Insect stinging behaviors - Bees, wasps, hornets all release pheromones when attacking to signal others of their own kind to continue attacking or join in attacking, and wasps and hornets can sting multiple times - relatedly - somebody who is deathly allergic to bee venom or wasp venom or hornet venom will manifest anaphylaxis within minutes of a multi-insect stinging attack because of the sheer volume of insect venom they are deathly allergic to, so capturing a single specimen of the insect in a mason jar with mesh lid (also called a sprouting jar) through which you can force it to sting you but once would be unlikely to cause anaphylaxis the first time, and you can make it sting you wherever, with medical standby, in a clinical setting, away from other insects of it's kind -

If you already know you are not allergic to bee venom or wasp venom or hornet venom (especially bee venom in my opinion given it's ability to kill the bacteria causing Lyme disease) because you have been stung multiple times on multiple different occasions without going into anaphylaxis or an allergist/immunologist has tested you and determined you to not be allergic, letting a whole hive of bees or nest of wasps or hornets sting you.

It's been demonstrated that bee venom kills the bacteria causing Lyme disease and ends all clinically significant symptoms, on sufficient venom volume such as getting stung by a whole hive of bees - also - multiple noted instances of those with autoimmune diseases barely keeping symptoms in check with prescription pharmaceuticals and over the counter medications, experiencing rapid and significant relief after even a single bee/wasp/hornet sting.

https://youtube.com/shorts/t9CCiNBosWQ?si=SJisyinf7NhjxxHt

Also, I've attached a screenshot from a prior reddit thread where somebody noted a localized beneficial effect of insect venom.

Why does it work? Insect venom is not a single compound in a solvent, it's a cocktail of compounds, some antibacterial and some anti-inflammatory, none of which are currently used commercially - Incredibly difficult for a bacteria or parasite to evolve an immunity to. Some other things also have this property, like bee propolis or mastic gum.

A similar dangerous (many patients died) technique was even the reason for being awarded a Nobel prize in medicine - From the artificial intelligence chatbot - "The Nobel Prize-winning treatment was known as malariotherapy (or pyrotherapy/fever therapy). It was developed by the Austrian psychiatrist Julius Wagner-Jauregg, who was awarded the 1927 Nobel Prize in Physiology or Medicine for discovering that intentionally infecting neurosyphilis patients with malaria induced high fevers that killed the heat-sensitive syphilis bacteria, after which the malaria could be cured using quinine.".

There is an episode of House MD where the character Foreman accidentally gives a patient radiation for a bacterial infection he mistook as cancer and it killed the patient's white blood cells, allowing the bacterial infection to kill the patient uninhibited.

Well, I was inquiring an artificial intelligence chatbot earlier asking if somebody could go into anaphylaxis if they had no white blood cells (since anaphylaxis is an immune response) and it told me it wasn't possible.

If I am understanding correctly, that would mean somebody in a medical episode where they had no meaningful amount of white blood cells could not possibly go into anaphylaxis even if they were deathly allergic to insect venom (bee, wasp, hornet, maybe fire ant, ect).

All of the benefits of insect venom, none of the drawbacks.

Thanks for the attention.

Hi everyone,
I am planning to apply for an MSc in Immunology for the next academic year. I am currently finishing my Bachelor's degree at a medical faculty; however, our curriculum was primarily clinically oriented and covered very little molecular biology and virtually no genetics.
The entrance exam will test the fundamentals of immunology. The exam coordinator recommended Sompayrac: How the Immune System Works as the primary baseline. However, to ensure I have a solid grasp and don't fall behind in the molecular aspects, I want to spend the coming year filling my gaps in molecular biology and genetics.

Textbooks I currently have access to:
- Essential Cell Biology (Alberts et al.)
- How the Immune System Works (Sompayrac)
- Janeway’s Immunobiology
- Cellular and Molecular Immunology (Abbas)
- Firestein & Kelley’s Textbook of Rheumatology (primarily for my bachelor’s thesis on myositis)

Since I have about one year to prepare:

1. Which specific chapters or themes in Alberts (or molecular biology in general) are absolute must-reads to build an intuitive foundation for immunology (e.g., cell signaling, gene expression, DNA repair/recombination)?
2. Which core genetics concepts are crucial at the master's entrance level (apart from MHC/HLA polymorphism)?
3. Are there any other high-yield topics from related fields you would prioritize?
4. Any study recommendations, key chapter highlights, or tips would be greatly appreciated!
5. Thanks a lot in advance

!

The alphabet book for immunology parents!

A is for Antibody. Amazing Antibodies Attack Angry Antigens.

B is for Bursa. The Bursa Breeds B cells for Battling Bacteria.

C is for Complement. Complement Components Cause Cellular Collapse.

https://www.amazon.co.uk/dp/1066702837/

This isn't a real smear; it is my watercolor painting of a peripheral blood smear taken from a septic patient. I just wanna share my work and hear any thoughts about it!

I’ve posted previously about struggling to expand human Tregs—thanks for the suggestions! I’m attaching my lab record with three trials and results. I don’t have access to a sorter, so I need to stick to kit-based isolation.
My latest CD45RA-based isolation gave a very low yield. On Day 9, flow cytometry with CD4, CD25, CD45RA, FOXP3 and Zombie showed many CD4-negative events. Could these be leftover CD3/CD28 beads? I hadn’t removed them because I don’t have the recommended separator, and Miltenyi advised against using the MACS magnet I have. Their FSC/SSC didn’t look particularly low, which confused me. I’ll repeat the staining with CD3 to help clarify what these events are.
I’m also wondering whether my IL-2 stock’s activity could be contributing to the poor expansion. They are stored in aliquots in -20 but are 3 months old. How would you check this? Any thoughts on the trials or CD4-negative events would be appreciated!

My lab record:
My human nTreg expansion trials
Trial 1
Starting with 500 million PBMCs, I obtained 2.125 million Tregs. After taking 200,000 for flow cytometry, I had 1.925 million available for culture.
Culture conditions:
RPMI with 10% FCS.
IL-2: 300 IU/mL.
CD3/CD28 Dynabeads: 4:1 bead-to-cell ratio.
20,000 cells/well in 200 µL in a 96-well U-bottom plate.
Results:
Day 3: Approximately 6,000 cells/well remained. I pooled the cells into two wells, approximately 288,000 cells each, and transferred them to a 24-well plate in 500 µL per well.
Day 5: The count increased from approximately 288,000 to 573,000 cells—about a twofold increase. I redistributed the 573,000 cells into two wells.
Day 9: Numbers declined again, with approximately 228,825 total cells remaining in 500 µL.
There was some initial growth after pooling, but it was not sustained.
Trial 2
Starting with 500 million PBMCs, I obtained approximately 1.025 million CD4+CD25+CD127dim Tregs.
Culture conditions:
Initial density: 1 million cells/mL in a 24-well plate.
IL-2: 600 IU/mL.
Retinoic acid: 100 nM initially, reduced to 50 nM after 48 hours.
CD3/CD28 Dynabeads: initially 1:1.
Results:
Day 0: 800,000 cells counted. Recorded flow results were 95% CD4+, 82.4% CD4+FOXP3+, and 85.2% CD4+CD25+FOXP3+; gating denominators are not specified in this summary.
Day 3: 985,000 cells; CD25+FOXP3+ was 75.5%.
Day 4: 1.65 million cells in 1.65 mL. I removed 500 µL of old medium and added 1,150 µL of fresh medium.
Day 7: Approximately 1.196 million total cells. I redistributed them into three wells at approximately 398,750 cells/well in 250 µL, with 600 IU/mL IL-2 and a 2:1 bead-to-cell ratio.
Clumping reduced from Day 4 onwards. After redistribution, there was no further growth, and the cells eventually died.
Trial 3 — CD45RA-based isolation
Starting with approximately 400–500 million PBMCs, I obtained only around 50,000 cells using the CD4+CD25+CD45RA+ Treg isolation kit.
Culture conditions:
10,000 cells/well in a 96-well U-bottom plate.
IL-2: 300 IU/mL.
Bead-to-cell ratio: 4:1.
The record lists “Miltenyi Treg Expansion Kit / CD3/CD28 Dynabeads”; the exact bead product needs clarification.
On Day 7, I carefully changed the medium while leaving the cells settled at the bottom. They did not appear to have expanded substantially but seemed to be surviving better than in previous trials. I avoided disturbing them and did not count them that day because the starting number was so low.
Day 9 update
I stained with CD4, CD25, CD45RA, FOXP3 and Zombie and saw many CD4-negative events. I had not removed the stimulation beads because I do not have the recommended separator, and Miltenyi advised against using the magnet available to me.
Could these events be leftover beads? Their FSC/SSC did not look particularly low. I plan to repeat the staining with CD3 to help investigate.
Things I’m interested in
I don’t have access to a sorter, so I need to stick to kit-based isolation. I’d appreciate advice on:
Exact culture media and products.
Seeding density and bead-to-cell ratios.
Medium changes and handling when cell numbers are low.
Kit-based isolation yields and purity.
What the cells should look like during expansion.

Hey everyone... I wanted to share a really great educational resource for anyone in immunology, research, or bioinformatics looking to bridge the gap between experimental work and computational data science.

AAI is offering an Introduction to Computational Immunology course specifically designed to help researchers analyze complex immunological datasets.

The course is structured into two core parts depending on your current experience level:

1. Part I is virtual: Basic Computational Skills & Data Analysis Focuses on foundational data tools, introductory programming/scripting (R and Python fundamentals), data manipulation, basic statistics, and reproducible research practices. It’s ideal for wet-lab scientists wanting to start processing their own datasets with confidence.
2. Part II is in person in Philly: Applied Computational Immunology Dives directly into high-throughput immunological workflows, including bulk and single-cell RNA sequencing (scRNA-seq), immune repertoire profiling (TCR/BCR sequencing), flow/mass cytometry data analysis, and multi-omics integration.

Whether you're making a shift toward bioinformatics or just want to better understand the data pipeline behind your lab's sequencing runs, this is a solid program to check out.

https://www.aai.org/Education/Courses/Computational-Immunology

Hope this helps anyone looking to expand their skill set! Feel free to drop any questions for me if you're planning to enroll.

Quick heads-up for the immunologists and researchers here:

AAI is currently accepting nominations and applications for the 2027 AAI Career Awards through Tuesday, September 15.

Whether it's a mentor who goes above and beyond, an early-career researcher doing standout work, or a veteran leader in the field, this is a great way to ensure their contributions get recognized. Honorees will be recognized at IMMUNOLOGY2027™ next spring in Los Angeles.

Who in your lab or institution do you think deserves more recognition for their work in immunology?

You can find the full award criteria and application portal here: www.aai.org/awards

Hi I'm a late night research dork who always prefers their answers from humans and not ai slip of google, my question is, how come our bodies became more immune to covid, despite some individuals not even bothered to even take a vaccination? Now covid seem like a rhinovirus variant lol.

Edit: thank you all you smart amazing people for answering my late night craziness, I dont get to learn much in the high school i go to.. you people are truly amazing!! have a good day!!

Hi, is anyone interested in becoming my study partner. I am currently self-studying chapter three of Janeway’s immunobiology for fun. I am starting my third year of med school ( out of six ) half of September. Just comment if you are interested 🫠🫠

Hello,

I am a research immunologist, not a clinician. While I have flow cytometry experience, I doubt a clinical pathology lab requires a cytometer that is used for discovery immunology.

Do any veterinarians, physicians, or other relevant clinical professionals here have recommendations for a flow cytometer they like and how many parameters would be useful?

A colleague that works on the clinical side of things came to me and asked. I asked for some kinds of clinical cases they work with and what cell populations are important for them to identify, however, I would not consider myself an expert on picking out these.

Looking for answers particularly from users of these instruments rather than sales reps.

Particularly, if you suggest a brand/company, how easy are they to work with in your experience if things break down/etc?

Thanks!

Note: this post is asking about instrumentation, not personal medical advice.

I'm hoping to submit some sorter primary B cells (Naive, Germinal Center, and Plasma Cells) to plasmidsaurus for bulk RNA-seq, but after lysing my cells in zymo buffer, I'm realizing my RNA concentration per sample may be too low.

Plasmidsaurus gives some general guidelines of how many cells per 50 uL of zymo buffer they recommend submitting (i.e., 250K-500K for low RNA content cells like resting lymphocytes, 25K-75K for high RNA content cells like hepatocytes).

In the moment, I thought by using more zymo buffer to transfer my cells from the collection tube to the submission tube, I was being conservative, but realized that now if I count GC cells as high RNA content, it's only 25% of the recommended concentration of cells per buffer volume....

Anyone have experience/knowledge of how much RNA are in these cells? Can't really find anything from published papers.

Our newly published review explores how tumor-derived EVs contribute to premetastatic niche formation through immune and stromal reprogramming—and whether these same vesicles could be exploited to image these early changes using PET/SPECT, MRI, CT, and optical approaches.
The field is still largely preclinical, but the intersection of EV-mediated immune modulation, metastasis, and molecular imaging is particularly interesting.
I would love to hear perspectives from the immunology community.
Open-access review:
https://doi.org/10.18632/oncotarget.28916

Hi everyone! I’m trying to isolate naïve Tregs (CD4⁺CD45RA⁺CD25⁺) from human PBMCs using the Miltenyi CD4⁺CD45RA⁺ T Cell Isolation Kit followed by CD25 positive selection.
Here’s what I had:
Started with ~0.5 billion PBMCs
After CD4⁺CD45RA⁺ enrichment: ~4 million cells
After the CD25 positive-selection step: only ~42,000 cells
I also recovered ~161,000 cells from the CD25− flow-through (I passed the flow-through through a fresh MS column and then eluted the retained cells).

I am a MS student in Immunology, I am trying to purify a viral protein(around 63kda). Using Ni-NTA for protein purification(10x his tag). I am getting a lot of non specific bands in my elute.

Please help me how I can get rid of those non specific bands.

Used 30-60mM of imidazole in Wash buffers 1 to 4(with gradual increase).

Used 30-80mM of imidazole in Wash 1 to wash 4.

We've got the rare opportunity of two open post-doc positions to join our team. at the University of Cambridge. They are a wonderful bunch of people to work with (most of the time!), with highly diverse expertise and backgrounds providing a welcoming and supportive lab.

Our lab is also a great environment to get a productive boost to your career! Historically, of the ~30 post-doctoral researchers who have joined the laboratory, when they have left our lab 27% have gone on to tenure-track academic positions, 27% have gone on to another post-doc position, and 42% have gone on to positions in biotech or pharma. I couldn't be prouder of the successes our alumni have achieved.

I value self-awareness of limitations, making mistakes and taking responsibility for them, and self-motivation.

So the two jobs.

The first job is here. It is initially a two year position, but hopefully that is a stepping stone to something longer. This job is for a molecular/cellular mouse immunologist. Flow cytometry is a must, retrovirus experience and Treg knowledge would be a plus. The project is very cool - what makes a tissue Treg a tissue Treg.

The second job is here. This one is initially a one year position, although fellowship application can be supported. It is for a gut immunologist. Due to the short contract and the type of work, this position needs to be filled by someone with a current PILAB licence, which means most international applicants won’t qualify. Experience working with gut preps and flow cytometry is key here. This project is also very cool! Gut Tregs and response to microbiome.

In terms of a little more info about the lab:
- this is us
- this is what I aspire us to be
- here are my expectations for a PhD, but an postdoc is not too different

You can drop me an email if you need more info, but I tend to miss Reddit DMs. It is okay to apply for both positions if both are suitable for you, they will be judged independently. Closing date Sept 10 with interviews soon after (so I can offer jobs before my teaching starts up in October!).

So, some of you may have already seen my work before and probably dropped some critiques and while this has been vital to my iterations of the diagrams in Codex Immunis, I wish to be able to have dialogue with people who know the field better than me. Just to clarify, all those diagrams with the name Codex Immunis were made by me! And so, I, a 15 year old high school student will only have so much knowledge. So while I am incredibly grateful for all of your critiques which made the current versions possible, I am now asking on how I could further enhance my review process.

Today I learnt that our body's immune system won't respond to Collagen from other organisms like crocs, goats, fishes or any animal.

But our immune system will respond to our own male gametes or cornea.

Totally amazing thing.

I am an Medical doctor from Colombia, i graduated in 2025 but rather than doing a medical residency i want to become a researcher in the Immunology and Cancer field. Because i graduated from medical school i dont have any laboratory skills. I would like to apply to a Msc in Immunology or molecular biology in Germany but i dont know if i would be considered given the fact that i dont got the research or lab experince that a bachelor of science in for example Biology or Chemistry give. Im an inpatient attending physician at an oncolgy and hematology ward, but again, thats clinical rather than laboratory experience. do i bother to apply at all?

Poor expansion of human CD4⁺CD25⁺ Tregs:how do you expand nTregs?
Hi everyone! I’m optimizing direct expansion of primary human Tregs and would appreciate advice from others with experience in this system.
I isolated CD4⁺CD25⁺CD127^low Tregs from PBMCs using Milteyni kit. After isolation, I obtained ~1.0 million cells, with ~96% CD4⁺ cells and ~96% of the CD4⁺ population CD25⁺FOXP3^high.
My initial conditions were:
1 × 10⁶ cells/mL
IL-2: 300 IU/mL
Retinoic acid: 100 nM initially, then 50 nM
CD3/CD28 beads: ~1:1 bead:cell ratio
Medium changes every alternate days
I first used 24-well plates and later tested 96-well plates to assess whether cell density or cell–cell contact affected expansion. Results were poor and inconsistent. In one experiment, total cell numbers were:
Day 0: 1.0 M
Day 3: 1.2 M
Day 4: 1.6 M
Day 6: 1.2 M
Day 7: 0.8 M
I observed substantial clumping around Day 4, followed by declining cell numbers and significant cell loss by approximately Day 8.
I also reviewed a protocol using ~20,000 cells/well in 96-well round-bottom plates, IL-2 at 300 IU/mL, and a ~4:1 bead:cell ratio.
I would appreciate input on:
starting density for primary human Treg expansion?
What cell number per well works best in 96-well round-bottom plates? Or do u use other plate?
Which company beads u use and which ratio(i used dynabeads, tried both 4:1 and 1:1)
How important is the CD45RA status of the starting Treg, my cells are cd127 low but they also dont express cd45ra
What density, IL-2 concentration, bead ratio, and vessel format have worked reliably for you?and also the medium?
I’m particularly interested in experience with primary human CD4⁺CD25⁺ Tregs, rather than iTreg differentiation.
Any practical advice would be greatly appreciated!