Science topic

Immune Response - Science topic

The platform for discussion on evolution, regulation and understanding the mechanism of immune response in Humans and other living organisms.
Questions related to Immune Response
  • asked a question related to Immune Response
Question
2 answers
Hi. I have a question of the MHCII expression in mammal. What I know so far is that the expression of MHCII is codominant, alleles from both mom and dad will be expressed. However, I would like to if the genotype of MHCII affect the phenotype. For example, would the MHCII wild type animals express more MHCII protein than the MHCII heterozygous animals? Also, would the MHCII WT animals process more antigen and have a stronger immune response than the MHCII heterozygous animals? Assuming only one specific type of antigen is processed in this case.
It will be very helpful if you can post some related literature. Thanks for your time!
  • asked a question related to Immune Response
Question
3 answers
I need information about general vegetable oil as adjuvants for my studies.
  • asked a question related to Immune Response
Question
7 answers
About our apper: The First Evidence of a Disturbed Immune Response toward Candida Albicans in Patients with Sars-Cov-2 and Co-Morbility. A Novel Case Report. Do you have a similar patients? Your experience?
Relevant answer
Answer
Thanks a lot for your response if you have an intesresting patient we can publish a case report
  • asked a question related to Immune Response
Question
15 answers
As we know the immune system in both children and adult is impaired. The question is that why the covid-virus infection is very rare in children compare to young and adult people
Relevant answer
Answer
Immunity level
  • asked a question related to Immune Response
Question
5 answers
Immune response against viral infection is cell mediated which releases many cytokines. It is clear that, in covid-19 infection level of cytokines is very high and called ' cytokine storm". Complications due to covid-19 infection is mostly due to this cytokines and can endanger the life by internal blood clotting. Other than remission of fever paracetamol has no anti-inflammatory action and this drug has no role on preventing release of cytokines. So use of paracetamol in this case will decrease fever escaping cytokines complications. On the other hand, along with action of decreasing fever NSAID can help to block formation of cytokines and thus prevents much of the complications due to covid-19 infection and decrease mortality. No doubt steroid is better than NSAID in this respect but many physician will hesitate to use steroid initially without confirmation. In all cases famotidine should replace PPI.
Relevant answer
Answer
Not at all . Paracetamol is safer than NSAID ( severe side effects and not to be used for ulcer , kidney pateints ) . HChQ is better than NSAID to reduce inflammation of Covid19 .
  • asked a question related to Immune Response
Question
8 answers
A very important study from Denmark published yesterday in the Lancet on 4 million people focused on reinfection of sars-cov-2.
The protection rate for those who have been infected is 80% for adult, and 47% for those > 65 in age.
It means there are 20% and 50% chance of reinfection in adult and older, respectively.
What could be the possible explanation? Immune response differences, CD4, B cells, and TH17 efficacy.
Do we have to reoptimize vaccine guidelines for whom have been previously infected?
Relevant answer
Answer
Immunosenescence as the root cause of older people at increased risk of COVID-19 infection, reinfection, outcome, and response to vaccination
It is well known that older people have immunosenescence that predisposes older adults not only to SARS-COV-2 infection, but also reinfection and poor outcomes. Even the vaccines have poor response in the older people. Strategies are being developed how to tackle Immunosenescence to reduce infection risk and improve response to COVID-19 vaccine (1-4).
  • asked a question related to Immune Response
Question
17 answers
What will be the consequence of the coronavirus vaccine on a patient’s health?
Relevant answer
Answer
  • asked a question related to Immune Response
Question
11 answers
Can someone conclude to me how we measure immune responses in Animal models in pre-clinical studies?
I know there are many ways and many essays. I want an opening to know how to start thinking in this part of immunology.
Thank you
Relevant answer
Answer
Hi (copy from your question into discussion loop ...)
If your aim is study immunogenicity of human drugs in animal models then best way is analysis of guidances by FDA/EMA :) and total ADA (anti drug antibodies), fraction of Nab's (neutralising - high affinity antibodies) and seroconversion time. Such data could be colected after sigle dose of human protein. Sometimes pilot study is useful. Sometimes best way to analyse immunogenicity of human drug in animal model is simply analysis of drug pharmacokinetics. Then you just see what the difference is with Nab's because this fraction of antibodies increases the clearance of the drug that is administered. You can measure the most important cytokines and cells. The meaning of such work increases when your model is relevant in pharmacokinetics and pharmacodynamic meaning. Please remember that using the animal models for comparision test versus reference drug immunogenicity could be valuable (see guidelines below) but translation into humans impossible and off value. Please remember that different doses of the duman drug can induce different immune responce. For example higher dose of the drug may give lower immune responce if drug has immunosupresive action. Moreover in case of some drugs multiple dosing may decrease immune responce for example ustekinumab or infliximab ....
Some explanations from old FDA guidance could be usefull in your work (document is not available now)
Animal immunogenicity assessments generally do not predict potential immunogenic responses to protein products in humans. However, when differences in manufacturing (e.g., impurities or excipients) between the proposed product and the reference product may result in differences in immunogenicity,
measurement of anti-protein antibody responses in animals may provide useful information relevant to patient safety. Additionally, significant differences in the immune response profile in inbred strains of mice, for example, may indicate that the proposed product and the reference product differ in one or more product attributes not captured by other analytical methods.”
GUIDANCE FOR INDUSTRY SCIENTIFIC CONSIDERATIONS IN DEMONSTRATING BIOSIMILARITY TO A REFERENCE PRODUCT FDA 2012
" A risk-based evaluation, focused on the mechanism of action of the therapeutic protein product as well as results of animal and in vitro evaluations should be performed to determine the need for collection of preand post-dose cytokine levels in the early phase of clinical development. In case of a clinical adverse event, such an evaluation may provide evidence to support the clinical diagnosis of cytokine release syndrome and help distinguish this entity from other acute drug reactions "
Guidance for Industry Immunogenicity Assessment for Therapeutic Protein Products
Immunogenicity in animal models is not predictive of immunogenicity in humans.
Assessment of immunogenicity in animals may be useful to interpret nonclinical toxicology and pharmacology data.
Immunogenicity in animal models may reveal potential antibody related toxicities that could be monitored in clinical trials.
May reveal immunogenicity differences between biosimilar and reference product
The immunogenicity of therapeutic proteins- what you don’t know can hurt YOU and the patient
" Therapeutic proteins show species differences in most cases. Thus, human proteins will be recognised as foreign proteins by animals. For this reason, the predictivity of non-clinical studies for evaluation of immunogenicity is considered low."
but .......
" the comparison of the antibody response to the reference product in an animal model may be part of the comparability exercise both for similar biological medicinal products "
" Qualitative or quantitative difference(s) of product-related variants (e.g. glycosylation patterns, charge variants) may affect biological functions of the biotechnology-derived protein and are expected to be evaluated by appropriate in vitro assays. These differences and impurities may have an effect on immunogenic potential and the potential to cause hypersensitivity. It is acknowledged that these effects are difficult to predict from animal studies and should be further assessed in clinical studies. "
Guideline on similar biological medicinal products containing biotechnology-derived proteins as active substance: non-clinical and clinical issues
Some nice papers about could be useful
Hope it's helpful
Best regards
Tomasz
  • asked a question related to Immune Response
Question
6 answers
Covid-19 vaccination in asymptomatic sufferers? Immune response treatment or vaccination?
Relevant answer
Answer
In my personal opinion, asymptomatic Covid patients have a strong immune system, fight the virus and do not need vaccinations.
  • asked a question related to Immune Response
Question
29 answers
Antigens are processed by specialized macrophages to produce complex protein-RNA complexes that eventually produce iRNA. When this iRNA is introduced to primary B cells that have never seen the antigen, they produce specific antibodies to that antigen. Th macrophage produced RNA is incorporated into the the genome of these B cells by reverse transcriptase that now become "memory cells" capable of a secondary response when confronted with the antigen they had never come into contact before. Reverse ttranslation in the macrophage is the best explanation for the production of such specific iRNA.
Relevant answer
Answer
Stanley Laham Your tritiated nucleic acid (radioactive) may provide the 1st clue. There are numerous hurdles to challenge the Central Dogma as it appears to be rock solid. Cracks with caveats are seen in the published literature with one publication discuss the potentiality using prion as an example.
  • asked a question related to Immune Response
Question
8 answers
I would like to design an in vitro assay in which I examine the reaction of T cells among PBMCs to the COVID-19. Due to safety issues, I can't directly culture the virus with PBMCs. On the other hand, for the stimulation of T cells and for induction of a specific response to an Ag, the presence of immunogenic antigens is sufficient. Do you suggest the same method?
What other methods are available to check the function of T lymphocytes against the virus in vitro?
Relevant answer
Answer
If you're just doing antigen recall... why do you need the whole virus? I mean technically, if you want ALL the potential viral antigens, just get an inactivated virus and use that.
Otherwise, there are a lot of recombinant proteins for SARS-CoV-2 and the other coronaviruses, and there are also overlapping peptides.
Now if you want to look at what T cell responses are during a natural infection and you don't have a BSL-3, there are psuedovirus systems out there that you can use by inserting your protein of interest.
  • asked a question related to Immune Response
Question
27 answers
Immune responses to infections with by a corona virus vary widely and are appear to be related to the development of most severe complication, acute respiratory distress syndrome. Since survival of patients respondingto the virus in this way depends on respirators support, mechanical ventilation and extracorporeal oxygenation, therapeutic methods which demand highly specialized medical and nursing staff, human resources which become scarce in an epidemic or pandemic. Since vaccination are not available in newly emerging corona virus epidemics it would be interesting to know if and which targeted pharmacological modulation of immune response early in the course of an infection could help to reduce the need for intensive care and/or improve the outcome of respiratory support.
Relevant answer
Answer
Detection of Immunoglobulin M (IgM), IgA, and IgG Norwalk Virus-Specific Antibodies by Indirect Enzyme-Linked Immunosorbent Assay With Baculovirus-Expressed Norwalk Virus Capsid Antigen in Adult Volunteers Challenged With Norwalk Virus
  • asked a question related to Immune Response
Question
2 answers
Greetings,  I am looking for your input on the following question: We are trying to design a study where we will inject sEVs (exosomes) isolated from human plasma and tissue biopsies to mice to look for some biological effects. However, there is a concern regarding the immune response that human sEVs might trigger in mice, and that this might impact the study outcomes.  Did anyone try something like that or perhaps see a published study doing something of that sort?
How would one minimize such a response if it does occur?  Thank you in advance for any advice!
Relevant answer
Answer
Hello Dmitri,
We have evaluated the safety and toxicity profile of human embryonic kidney-derived EVs after intravenous administration in mice. Overall, we did not see toxicity effects or immunogenicity associated with our EV treatment.
Similar results were also reported by other groups (here is a nice study).
However, many factors including the EV dose, source of the cells, frequency of EV administration, administration route, etc. can have an impact on the overall toxicity of EVs. I would recommend you to perform a small safety study before your experiments to be sure that your EVs do not induce toxicity and immunogenicity in vivo. I hope this helps.
Best wishes,
Elisa
  • asked a question related to Immune Response
Question
6 answers
Vaginal abscess in pregnant women caused by Staphylococcus aureus and Trichomonas vaginalis is one of the most important disease, this infection is still medically uncontrolled because of poor health awareness due to the absence of personal hygiene in some women.
  • asked a question related to Immune Response
Question
9 answers
I'm getting zero or no ct value in my gene expression analysis during RT-PCR analysis. I'm not understanding the meaning of zero ct value, how I should interpret the data now. Should I consider this ct value for further analysis of gene expression.
Relevant answer
Answer
Hello Sandeep, how are you? Well, there are some reasons for this trouble. Primary, maybe your primers are not working or your cDNA samples are not well synthesized. As answered before, you can perform a conventional PCR to check if the primers are working. Also, use your endogenous control (GAPDH, ACTIN, 18s) and one sample that you know that your primers work to verify if your samples are well synthesized. Run an agarose gel and check the size of the fragment. If the gel does not appear any band, your sample are compromised or your primers are not working. Another problem is that your sample has a low quantity of your target gene, and the amplification curve is not detected by the machine even after several cycles. Usually we work with a diluted cDNA sample. The thermocycler are very sensitive to a low quantity of copies, but there is a limit to that. Maybe you need to concentrate your sample to initiate the amplification earlier. Thus, the thermocycler can detect the amplification signal. After all this, if you still detecting no CT value of your target gene, but your endogenous control is good, maybe you should consider that your target gene do not express in your sample. Good luck.
  • asked a question related to Immune Response
Question
21 answers
NK cells or interferon alpha? 
Relevant answer
Answer
The immune response that develops in a viral infection is first given by the innate immune response. The virus enters the cell and deposits its genetic material that is recognized by endosomal TLRs (TLR3, 7, 8, 9) or by proteins of intracytoplasmic recognition (MDA-5, RIG-1 and DAI) of foreign DNA and RNA, these proteins recognizing foreign nucleic acid will activate the kinase proteins that will then activate the transcription of IRF (interferon regulation factor, IRF1,3 and 7 ) that enters the cell nucleus and activates the transcription of interferon type I (alpha and beta). This cell will produce type I interferon that comes into contact with neighboring uninfected cells by interferon I and II receptors, getting the cell into an antiviral state. This state is caused by the activation of the transmission pathways of jack-stat signals that induce the expression of genes whose products interfere with the replication of the virus in uninfected cells. We must remember that type I interferon (alpha and beta) increases MCH-I expression, activates NK cells and promotes differentiation of Th virgin lymphocytes to Th1.
I hope the information provided helps you, regards.
Español
La respuesta inmune que se desarrolla en una infección viral se da primeramente por la respuesta inmune innata. El virus ingresa a la célula y deposita su material genético que es reconocido por los TLR endosomicos (TLR3, 7, 8, 9) o por las proteínas de reconocimiento intracitoplasmatico (MDA-5, RIG-1 y DAI) de ADN y ARN extraño, estas proteínas al reconocer ácido nucleico extraño activaran las proteínas cinasas que luego activaran la transcripción de IRF (factor de regulación del interferón, IRF1,3 y 7) que ingresara al núcleo celular y activara la transcripción de interferón de tipo I (alfa y beta). Esta célula producirá interferón de tipo I que entrará en contacto con las células vecinas no infectadas mediante los receptores de interferón I y II, consiguiendo que la célula entre en un estado antivírico. Este estado se da por la activación de las vías de transmisión de señales jack-stat que inducen la expresión de genes cuyos productos interfieren en la replicación del virus en las células no infectadas. Debemos recordar que el interferón de tipo I (alfa y beta) incrementa la expresión de MCH-I, activan las células NK y promueven la diferenciación de los linfocitos Th vírgenes a Th1.
  • asked a question related to Immune Response
Question
4 answers
need suggestion if someone can help in selection of most suitable but less costly and cheapest adjuvant to produce humoral response in mice animal model? Due to funds issue i just cant manage to opt for available adjuvants in market, so i just wonder if there is any alternate option for anything to be used as adjuvant in mice animal model experiment?
Relevant answer
Answer
Aluminum based adjuvants (aluminum hydroxide and aluminum phosphate) are powerful adjuvants in rodents, including mice, for protein antigens to elicit a high humoral immune response (antibodies). Selection of aluminum hydroxide or aluminum phosphate depends upon the pI of the protein. These adjuvants are cheap and you could prepare in-house. Please find attached an article describing methods to prepare these adjuvants in the lab and even in commercial scale. This article also discusses about selection of aluminum hydroxide or aluminum phosphate based on pI of the protein.
  • asked a question related to Immune Response
Question
3 answers
I am very new to the field of immunology and i am hoping to profit from the experienced scientist on research gate. I am working with a fibroblast cell line (RPE1 hTERT). Recently, i performed RNA sequencing and observed that genes involved in MHC 2 and antigen presentation are often down regulated when compared to my controls. I would like to validate my transcriptome data and then perform some functional assays to check if the antigen processing and presentation is working as efficiently as in controls. But i am not entirely sure about the assays that can be used to address my question. I would be thankful if someone could provide some suggestions. thank you.
Relevant answer
Answer
It's the clonal selection theory of adaptive immunity, meaning that only T cells with a TCR specific for the peptide presented via the MHCII can get activated and will proliferate (if the get the correct co-stimulatory signal at the same time). If you know, which MHCII alleles your cell line expresses, you might have a chance to know, which peptide they could theoretically present and then if you have a T cell line with a TCR specific for that peptide you can set up straightforward co-stimulation assays that are outlined in the paper. If you RNA-seq data is good, you don't need to phenotype your cells for their HLA alleles as you (or a good bioinformatician) should be able to parse out, which alleles they express right away. Then you can take it from there and follow the assays outlined in the paper from Ohyama et al.
All the best & good luck with your experiments,
Michael
  • asked a question related to Immune Response
Question
4 answers
Hello RG community,
I have a question regarding introducing exogenous T-cells into a nude athymic BALB/c FoxN1nu mouse model. I am studying the immune system's role in cancer dormancy (breast cancer). I am thinking about transfering mouse CD4+/CD8+ T cells into the mice (so that any response attributed to T-cells is from the transferred cells)
What would I have to take into consideration before performing such a procedure? Where can I get these cells from? And are there any protocols out there for such a procedure?
Thanks!
Relevant answer
Answer
I think, the most important rule should be coincedence of genetic backgrounds for recipient, tumor and injected cells. They should derive from BALB/c mice. Protocols can be very different in dependence of therapeutic range of injected lymphocytes, the aggressiveness of the tumor, etc.
  • asked a question related to Immune Response
Question
1 answer
Creo firmemente que este agente L, es muy evolucionado y usa la Ri del H para poder sobrevivir y multiplicarse y parte de las reacciones patologicas las crea la propia respuesta,desviada por el parasito!!!
Por que no le impedimos eso?
Quizas podamos hablar! Por correo: gsierra6352 @gmail.com
Gustavo
Relevant answer
Answer
Please make your question in English!
  • asked a question related to Immune Response
Question
3 answers
We are looking into the response induced by two plasmids (a backbone and a construction) and we wondered whether immune response is related to the quantity of mass (in bp) or related to the number of plasmids inserted into the cell (regardless their size).
Relevant answer
Answer
Hi again,
3µg for empty plasmid and 6µg for the construct.
  • asked a question related to Immune Response
Question
4 answers
Is there anyone who has information in regards to ELISA analysis done on crocodilians? Looking to assess parasite load and immune response in crocs, but I can only find one article who's tried this semi-successfully.
Is there a other/better way to measure immune response, or the lack of, in crocodiles in response to parasite load?
Cheers
Joe
Relevant answer
Hi Dr. Al-Salhie, unfortunately your link shows an equipment that is called "Crocodile" ( the commercial name of the instrument is crocodile, a device to pipette, wash and incubate plates only). However, Berthold technologies does not sell ELISA assays directed to crocodilian proteins, just as nobody else does for reptiles. Believe me, I searched everywhere for antibodies against snake proteins.
Thank you though, for a split second my hopes were high...
  • asked a question related to Immune Response
Question
4 answers
How do I identify the antigenic proteins or substances of an infectious organism which are involved in illiciting a particular immune response?
Relevant answer
Answer
See this link may be helpful
Regards
  • asked a question related to Immune Response
Question
10 answers
Hi everyone,
I'm working with immune response in colon cancer patients. Now I just wonder about that can I measure the NK cell activity? 
Thank you in advance
Da
Relevant answer
Answer
See this link of article is useful
Good luck
  • asked a question related to Immune Response
Question
2 answers
I'm performing some immunofluorescence for SPARC (osteonectin) in PMA differentiated U937 cells. I am wondering if anyone has looked at SPARC expression in macrophages and U937s in particular, and/or is aware of any literature on this topic. I haven't been able to find much published data, but maybe my search strategies are sub-par.
Relevant answer
Answer
agree with dr Torsten Goldmann
  • asked a question related to Immune Response
Question
5 answers
Recently, many reports have shown the adjuvant effect of ZnO nanoparticles or nanowires.
Some are showing good immunotherapeutic effects also. I am curious if anyone can help me to find out how ZnO is assisting the immune system.
I mean is there any specific mechanism?
Relevant answer
Answer
Hello
This is an interesting topic.
Zn is useful in the body in very small amounts and ZnO helps to enrich the blood with oxygen. This makes it possible to enrich the regeneration of red blood cells. The remaining Zn is excreted through the syringe system or accumulates in the bones if it is in increased doses.
  • asked a question related to Immune Response
Question
2 answers
 Which type of NAE in moss? 
Relevant answer
Answer
Guy Dutau,  thank you for your response
  • asked a question related to Immune Response
Question
2 answers
did it induce immune response or cause side effect independent of the cargo?
Relevant answer
Answer
works fine, but everything depends on the exosome dose (make sure its physiological) and purity (make sure there are no admixtures in your prep- eg trace amounts of chemicals)
  • asked a question related to Immune Response
Question
4 answers
Hi there! I am looking to characterize the immune response in an in vivo murine model of transplantation. I will be analyzing the spleen to assess for the presence and activation states of T cells, B cells, and Macrophages. 
What markers should I use to reliably differentiate each cell type and its activation state when utilizing flow cytometry. 
Any additional comments to help characterise the immune response are welcome!
Thanks
Relevant answer
Answer
Hi Keerit,
here are some suggestions.
For T cell identification (CD3 or TCRb plus CD4 and CD8); subsequently activation markers include: CD44 (upregulated during persistent T cells activation), CD62L (naive T cell marker that's inversely downregulated), CD69 (upregulated intermittent T cell activation), and possibly CD25 (traditionally a Treg marker, but also expressed on activated T cells)
For B cell identification (CD19 and/or B220); subsequently activation markers include: CD54 (ICAM1), MHCII, CD80, CD86, surface IgG
For proinflammatory Macrophage identification (CD11b and/or F4/80); subsequently activation markers include: MHCII, ICAM1, iNOS and NADPH oxidase subunits like i.e. p47phox
Most of the above mentioned markers are on the surface and therefore detectable with a simple extracellular staining. You can of course add, intracellular cytokine/ transcription factor stainings for TH1/ TH2/ TH17/ Treg differentiation. I'd particularly have a look at TH1 and Treg stains for example with IFNg and FOXP3. Moreover, you can give a thought to a perforin and granzyme B staining on T cells, which may particularily be suitable for extrapolating the likelihood of transplant rejection.
Hope that helps!
Good luck with your experiment!
Michael
  • asked a question related to Immune Response
Question
3 answers
How long can I keep HK-bacteria, and at what temperature? Previous protocols have stored it at 4C for up to 4 days, but ideally I'd like to be able to store it for longer in order to make bulk batches and aliquot it. I'm only trying to induce an immune response in caterpillars, without having to worry about pathogenic toxicity. Or are there better alternatives for inducing immune responses, such as using LPS vs. whole bacteria?
Relevant answer
Answer
Heat-killed bacteria can be stored at -20.
Compare to purified bacterial ligands (LPS, PAM3CSK3, etc) you can expect more variations in your experiments using HK bacts. So if you want to have a robust stimulations you are better of using specific bacterial (TLR) ligands.
Try invivogen website :)
  • asked a question related to Immune Response
Question
10 answers
Hello,
I am undertaking a research project recruiting patients, taking a blood sample, culturing then stimulating monocyte-derived-macrophages and then measuring their immune response (e.g. cytokine levels) following the application of various stimuli. I plan to recruit healthy matched controls from the same geographic area but am a little confused as to how much I should match these controls.
I plan to knock on randomly selected households in the same neighbourhood of the case and invite healthy controls that fit the matched criteria to take part in the study but don't want to be too constrictive both for statistical and logistical reasons, and risk over-matching.
If sex and age matched controls is advised what is the most practical way of finding, for example, the age range that I wish my matched control to be in. Say I have a male patient born 12/2/75 (fictitious patient of course), do I calculate to create an age range that spans e.g. 4 years over this DOB - that is 2 years either side of the DOB? (11-Feb-73 to 11-Feb-77). This seems fiddly but obviously more accurate than looking for a control between 40 - 44 years of age.
Any help much appreciated.
Relevant answer
Answer
Interesting topic.  As is typical, the answer will depend on your research question.  It isn't clear to me whether the cases in your study are of TB disease or latent TB, and if they are co-infected with helminths.  The strongest risk factor for all of these is probably foreign birth, so if you select controls from the general population you will likely find this to be the strongest association in your study.  So you may want to match on that if it is not a variable of interest.  Age is less strong for TB but still important so matching might be important.    
If you are interested in studying whether factors, such as co-infection, might lead to progression of latent TB to TB disease, then you might want to select controls who have LTBI.  This would probably obviate the need for matching on foreign birth or age.
If you simply want to see the differences in immune responses between co-infected vs mono-infected, you might want to compare 4 groups: uninfected for both, infected for both, and mono-infected TB and helminth.  In that case, I would think that matching would be related to those variables which are known to affect your immunological outcomes of interest, which will depend on which outcome you select.  I would think about age and foreign birth as potentially important sources of confounding, but also comorbidities and other factors.
  • asked a question related to Immune Response
Question
3 answers
I have tried coating plates with OVA  using carbonate buffer. Whilst the first well may give a high OD read-out the samples do not result in a titration curve
Relevant answer
Answer
Dear Donna, yes, the protocols are very similar. I titrate serum IgGs, and do not have experience with your sample types. Nevertheless: Some considerations that might give you a clue about the source of your problems:
- How do you prepare the serial dilution of your samples? Do you use a too high dilution factor that might be responsible for the sudden signal drop?
- Do you observe the same problem for all of your sample types (caecum, colon and faecal samples)?
- Do you have a positive control to check whether the current protocol works for IgA?
- Did you include a negative control to exclude other problems?
- Have you tried higher BSA concentrations for blocking to decrease unspecific binding?
  • asked a question related to Immune Response
Question
5 answers
Is there any way to detect/measure activation of TLRs (preferably specific TLRs) in tissue sections?
Relevant answer
Answer
Thanks for the additional comments! Unfortunately I don't think these approaches would be specific enough, because I'm talking about tissue sections and the mentioned downstream signaling molecules are not exclusively used by TLRs, so their activation doesn't prove TLR activation.
  • asked a question related to Immune Response
Question
5 answers
There are many type of carrier but we use the Keyhole Limpet Hemocyanin (KLH) is used in immunotherapy treatment for the emerging diseases. Why we use keyhole limpet hemocyanin as a carrier only? 
Relevant answer
Answer
KLH is a transporter that allows the immune system to function when the molecule of interest with which it is associated is too small.
This is particularly the case with most synthetic peptides which possess too few distinct epitopes to be taken up by dendritic cells on the one hand, then CD4 T cells and finally by memory B cells. It takes two to three separate epitopes to have a good immune response. KHL will then lend one or two epitopes to ensure an effective immune response.
In addition, KLH can be used to purify the target-KLH chimera, and use the residual anti-KLH response to follow the immune response.
The other carriers that can be used are either immunostimulants (Al3 +, TLR...) Or presentation particles (VLP, micelles, liposomes, nanoparticles, etc.)
  • asked a question related to Immune Response
Question
7 answers
Would attaching a carrier molecule help in enhancing the immune response? Also how do I ensure that antibodies are not directed against the carrier molecule? Are there any such carrier molecules that can serve this purpose?
Relevant answer
Answer
you may couple your peptide to a carrier protein such as ovalbumin or KLH, but take care to end up with a balanced molar ratio of peptide to carrier, for example 2 or 3 peptide molecules per carrier molecule
  • asked a question related to Immune Response
Question
5 answers
Hi all, 
Goin thru the online papers that describe immunisation and challenge experiments normally draws to a conclusion that the immunisation gives protection or even partial protection to the host. Are there any papers that showed no protection after the immunisation, even though the immunisation experiments are able to induce immune responses in the host? Thank you.
Relevant answer
Answer
In reality there are only articles that tend to show that seroconversion goes hand in hand with protection.
But the great majority of the works carried out all over the world lead to the opposite: an immune response giving no protection.
But these works are never published, without interest! What's the point!
For example: Influenza, HIV, Dengue, CMV, Leptospira, Pseudomonas, cancer cells ....
  • asked a question related to Immune Response
Question
6 answers
Why conjugated molecules create stronger immune response? Example protein-polysaccharide better than only protein or only polysaccharide? Is this time related?
Thank you.
Relevant answer
Answer
I suggest that you read a textbook chapter  on hapten -carriers and why they are important for the interaction of T and B cells!
  • asked a question related to Immune Response
Question
3 answers
We suspect that the 'ferocious' immune response that we observe is due to the presence of a conjugated ligand.
Relevant answer
Answer
Hello,
Fixing the cells, immunofluorescent staining with specific antibody and fluorescence microscopy. The protocol optimized for your cells and protein of interest is very important, it should be searchable.
  • asked a question related to Immune Response
Question
6 answers
I want to evaluate the humoral immune response to adenovirus vector vaccine carrying the HA gene of A/HongKong/156/1997/H5N1. I am working in BSL2+ lab,  But I don't have this virus in my stock. I evaluated the cell mediated immune response but, I want also to evaluate the humoral immunity.
Do you have any suggestions?
Relevant answer
Answer
Protein Sciences sells rHA from several viruses, including HK/156/97.  http://www.proteinsciences.com/
  • asked a question related to Immune Response
Question
1 answer
What`s the role of tissue-resident memory T cells in the mucosal immune response?
Relevant answer
Answer
Hi Qingrong
Look PDF attached may help you.
Good luck
  • asked a question related to Immune Response
Question
3 answers
Dear all,
Would you like to suggest me where I can get LUVA cell stocks?
Thank you very much,
Duy
Relevant answer
Answer
They are handled by Kerafast.  If you check their website, they are available.
  • asked a question related to Immune Response
Question
3 answers
This is an idea I have for my science research project. I have read through many articles regarding the potential effects of antibiotic resistance marker genes in GMOs on immune response, but I have gotten a mix of results. Half of the articles state that there is no danger (thus, no point in further researching it) while the other half states that there is a possibility. Would it be a plausible idea to research this topic? (Note: The articles were all a mix from before and after 2000, so I couldn't rely on how recent each experiment had been conducted) I would GREATLY appreciate it if anyone could direct me to articles that would help me decide whether to continue with this idea or not. Also, I have seen websites talk about the successful removal of ARMs from GMOs, but I have not found any research papers, so I cannot confirm this.
Relevant answer
Answer
Your question is very broad. To give you some hints: it all depends on the GMO and which antibiotic you are interested in.
  • asked a question related to Immune Response
Question
8 answers
A neutralising antibody is one, which in an in-vitro neutralisation assay reduces the viral infection in its presence. This is because it hinders in the infection process. (correct me if wrong)
On the other hand, can non-neutralising antibodies help at all in decreasing the viral load  in-vivo? They if not stop infection into the cell, can help in opsonisation leading to phagocytosis (or maybe by other mechanisms). Is this a process considered inferior to viral neutralisation? Does opsonisation help in viral infections (viruses generally intend to be endocytosed)?
Regarding antigen choice for making vaccines, antigens against which neutralising antibodies are made are chosen. If there is a fall in viral load with non-neutralising antibodies (in-vivo), can they too be used as vaccination targets?
Relevant answer
Answer
ADCC would work if you had an antibody against a component of the virus that shows up on the membrane of the infected cell.  but would not work (that i know of) if the antibody were against a molecule exposed on the virion, but not on the cell.  still.......in a mixed antibody response, there should be some antibodies that work via NK cells and ADCC.
  • asked a question related to Immune Response
Question
4 answers
I have HLA-A, -B and -C alleles and frequency data and wish to do multiple comparisons
Relevant answer
Answer
Dear Nicholas Nii-Trebi,
Please have a closer look to:
Best regards,
Thomas
  • asked a question related to Immune Response
Question
2 answers
I am focusing on the genes involved in the immune response (specifically for the generation of adaptive immunity)
Relevant answer
Answer
If you are looking to do RNA-sequencing analysis to get gene expression profiling, but do not have experience yet with bioinformatics, I would suggest using the Galaxy server:
You can find some tutorials here: https://github.com/nekrut/galaxy/wiki
-Benjamin
  • asked a question related to Immune Response
Question
3 answers
I am looking for a collaborator in Indonesia who is interested in immune responses to viral respiratory infections and looking at intervention trials in the area of antioxidants and reducing effects on childhood lung disease.
Relevant answer
Answer
Dear Collegue
I have colleague ... Dr  Rina Triasih Indonesian Pediatricians from Universitas Gadjah Mada .. PhD graduate from Melbourne University and published many international publication. If you need more information about her I will give you further contact information
Best wishes
Awalia Febriana 
  • asked a question related to Immune Response
Question
1 answer
We are working on the development of diagnostic systems for cystic fibrosis. We are having problems in the storage of our IRT proteins. The protein is degraded upon freeze thawing. Storage in PBS seems to disrupt the structure. We tried storage at pH:4.8. It works better, but we are still not sure what is the best method for storage. Any suggestion will help while we are pursuing our optimization work.
  • asked a question related to Immune Response
Question
7 answers
Hello everybody,
I am trying to produce ShRNA stable cell on RAW264.7. I succeed to get cells which is around 70-80% knockdown in puromycin selection condition. But the thing is when I measure the immune response, its totally opposite to my previous data and to the data which produce by SiRNA method (ShRNA cause increase immune response and  SiRNA cause decrease)
I have tried to produce shRNA stable cells three time, only 1 time I got the same data but this data is repeated only 2 times. 
Overall, there are no significant differences in cell condition, cell morphology or anything else. Does anyone have experience those kind of effects? would you please give me some advice? 
 Thank you very much
Relevant answer
Answer
it would help to know what you're stimulating and measuring as "immune response".  in the meantime here is a possibility.  when you use SiRNA, you're likely using something like lipofectin to get it into the cell?  that perturbs the membrane and perhaps stimulates the inflammasome.  so the cell gets at least partially activated.  if you're comparing the SiRNA to an empty vector, then the empty vector is also causing partial activation and the differences between control and experimental will be small.  when you use ShRNA, you wait for much longer to get a stable line, so the membrane perturbation will have settled down and your cells will be more in a "resting" state.
in any case, could you offer some more detail as to what you're doing?
  • asked a question related to Immune Response
Question
5 answers
 A 40 yr old immunocompetent host , with a short 3 day history of fever , breathlessness , deteriorated  rapidly and  died.
Non Smoker
Profession : Soldering work.
Microscopy Findings:
Septate hyphae with acute angle  branching  were seen in sputum, BAL and endobronchial biopsy . 
Bone Marrow Biopsy : No hidden malignancies . 
Relevant answer
Answer
  • asked a question related to Immune Response
Question
4 answers
Have come across a strange lagged response to an infectious agent where females seem to respond far more rapidly than males.
Is there anything in the literature regarding profound differences in immune response between the two genders or anything about gender and immune lags or immune feedback loops?
Relevant answer
Answer
Many thanks Willem. After some more analysis it looks like the time lags may also have something to do with different routes to infection between males and females, however, the differential immune responses are of vital importance.
  • asked a question related to Immune Response
Question
10 answers
There are many literatures that HIV can increase the risk of getting HCV infection. But what about the opposite direction? I have't got any information about that.
For people with HCV infection, do they have higher risk of getting HIV?
Relevant answer
Answer
Dear sir,
See this link may be useful for you.
Hepatitis C coinfection enhances sensitization of CD4(+) T ...
de C Körner - ‎2011 - ‎Cité 18 fois - ‎Autres articles... increased rates of CD4(+) T-cell apoptosis in HCV-infected HIV-positive patients ... Susceptibility to FasL-induced apoptosis was analysed by incubating isolated ... CONCLUSIONS: Our findings suggest a synergistic mechanism in HIV/HCV ...
  • asked a question related to Immune Response
Question
3 answers
Most researchers used different types of knockout mice for studying the immune stimulatory effect of certain chemical/extracts. Is there any difference in using the normal or knockout mice model for such kinds of study?
Relevant answer
Answer
Well the whole point of using k.o. mice to see the phenotype of a specific gene on different biological situations. It is higly depends on the qestion you ask. There are some specific models of spontanious desease development due to gene k.o. yet again you shuld consdider what is the scientific qestion you adressing with this. 
  • asked a question related to Immune Response
Question
8 answers
I need to study the T regulatory cells in pregnancy mouse model of malaria...which mouse strain I should use from the above two strains?
Relevant answer
Answer
Thank you Sir Yuksel...but why Balb/c any specific reason?
  • asked a question related to Immune Response
Question
2 answers
It has been demonstrated that IFN-gamma generation is related to the induction of protective immunity against Leishmania parasites following natural infection or vaccination, but this response is also known to be insufficient for inducing protection despite its main role. I would like to know why the cytokine response is not associated with a protective immunity. If anyone has experienced that, I would be pleased to share his/her experiences.
Relevant answer
Take a look at this review, it might help you.
  • asked a question related to Immune Response
Question
1 answer
Hi everyone!!...
I work with outer membrane vesicles (OMVs) for to evaluate the immune response in J774A.1 macrophages-like. The OMVs are suspended in PBS and I diluted in RMPI for test in the cells. Is necessary to heat the OMVs or there is some special consideration for this?
Thanks :)
Relevant answer
Answer
Dear Andres,
The following covers the answer to your question:
FEMS Immunol Med Microbiol. Author manuscript; available in PMC 2015 May 11.
Published in final edited form as:
FEMS Immunol Med Microbiol. 2012 Dec; 66(3): 436–444.
doi: 10.1111/1574-695X.12010
PMCID: PMC4426994
NIHMSID: NIHMS413557
Pluronic P85 enhances the efficacy of outer membrane vesicles as a subunit vaccine against Brucella melitensis challenge in mice
Neeta Jain-Gupta,1 Araceli Contreras-Rodriguez,2 Ramesh Vemulapalli,3 Sharon G. Witonsky,4 Stephen M. Boyle,1 and Nammalwar Sriranganathan1,*
Abstract
Brucellosis is the most common zoonotic disease worldwide and there is no vaccine for human use. Brucella melitensis Rev1, a live attenuated strain, is the commercial vaccine for small ruminants to prevent B. melitensis infections but has been associated with abortions in animals. Moreover, strain Rev1 is known to cause disease in humans and cannot be used for human vaccination. Outer membrane vesicles (OMVs) obtained from B. melitensis have been shown to provide protection similar to strain Rev1 in mice against B. melitensis challenge. In the present work we tested the efficacy of Pluronic P85 as an adjuvant to enhance the efficacy of Brucella OMVs as a vaccine. P85 enhanced the in vitro secretion of TNF-α by macrophages induced with OMVs and P85. Further, P85 enhanced the protection provided by OMVs against B. melitensischallenge. This enhanced protection was associated with higher total IgG antibody production but not increased IFN-γ or IL-4 cytokine levels. Moreover, P85 alone provided significantly better clearance of B. melitensis compared to saline vaccinated mice. Further studies are warranted to find the mechanism of action of P85 that provides non-specific protection and enhances the efficacy of OMVs as a vaccine against B. melitensis.
Materials and Methods
OMV Purification and characterization
OMVs were obtained from B. melitensis 16M as previously described (Gamazo, et al., 1989) but with the following modifications. Briefly, stock cultures of B. melitensis 16M were streaked on tryptic soy agar (TSA) plates and incubated for 48 h. Using cotton swabs, the bacterial culture was transferred to TSA slants containing 0.7% yeast extract and allowed to grow for 48 h. Next, the bacterial culture was scraped from slants using tryptic soy broth (TSB) (1mL/slant) and collected and spread on TSA plates containing 0.7% yeast extract. After an incubation of 48 h, Brucella cultures were collected with 200 µL phosphate buffered saline (PBS)/plate using a sterile cell scraper. The bacterial suspension was centrifuged at 15,191 × g and supernatant was collected. The supernatant was filter sterilized twice using 0.22 µm filters and the sterility of the filtrate was tested by inoculating 50µl into 10mL TSB and incubating at 37°C for 48–72 h. Absence of any bacterial growth confirmed the sterility of the filtrate. To obtain the OMVs, the filtrate was ultracentrifuge at 176,508 × g for 2 h. The pellet was washed in 50 mL PBS and harvested by centrifugation as above. The washing step was repeated twice and the pellet containing OMVs was resuspended in 250 µL PBS and stored at −20°C.
Concentration of protein in OMVs was determined using Pierce BCA protein Assay kit (Thermo Scientific) according to the manufacturer’s protocol. For the assessment of structure of OMVs by electron microscopy, OMVs were mixed with 2% aqueous uranyl acetate solution for negative staining and placed onto 200-mesh formvar, carbon coated copper grids (Electron Microscopy Sciences). Excess liquid was soaked away using filter paper and the samples were viewed at 80,000× magnification on a Zeiss 10CA Transmission Electron Microscope (Virginia-Maryland Regional College of Veterinary Medicine). To determine the protein profile, first OMVs were mixed with different concentrations of P85, dissolved in phosphate buffered saline, using a double hub-syringe and then mixed with 2× Laemmli buffer (Bio-Rad) containing β-mercaptoethanol and heated for 5 min at 95°C. The samples were electrophoresed on 10% SDS-PAGE gels (Invitrogen) and stained with Coomassie blue to visualize protein bands.
To view the full paper, please see attached file.
Hoping this will be helpful,
Rafik
  • asked a question related to Immune Response
Question
6 answers
I've been trying to find published studies about the physiological effects of saline injections in insects. So far, I have only found one paper (Jarosz, J (1988) Cytobios 53:19-29) that suggests saline injections increase levels of lysozyme activity and resistance to parasites in insects. I would like to know if anyone knows of more recent papers about this topic. I would also like to know why these saline injections are use as 'control treatments' for immunological assays if they seem to have an effect in immune response. Thanks in advance for your input!
Relevant answer
Answer
Correct! 75% survival
  • asked a question related to Immune Response
Question
12 answers
Hello,
I am going to work with a helminth crude antigen, so need a suitable adjuvant to appropriately elicit immune response in C57 black mice.
Thank you.
Relevant answer
Answer
Hi, just a short thought: Helminths usually initiates a Th2 type immune response. The C57Bl/6 strain is not a very good Th2 responder strain in comparison to other inbred strains such as Balb/cJ or NIH/Ola, which I have also worked with in allergy studies. (Allergy is also Th2 response). For example - I used the Th2-promoting adjuvant aluminiumhydroxide, but the antibody response was much higher in the two latter strains compared to the C57Bl/6-strain. So as Frederic Beaudoin points to, it is very important to consider the type of immune response you want to study and from this select strain and adjuvant. Best regards, Jitka
  • asked a question related to Immune Response
Question
10 answers
I want to study the result of the immune response which happens when we add certain number of cells to a foreign blood, like that which happens after kidney or bone marrow transplantation.
I need a usable protocol.
Your help is appreciated.?
Relevant answer
Answer
you can't.  the immune response in vivo will be the result of all sorts of signals that don't occur in vitro.  so it really depends on what you want to know.  if the strength of the response is all you need, then, as listed above, the mixed lymphocyte culture can give you a reasonable idea.  but if you want to know more than that (like what kind of response occurs to different transplants, or the kinetics of the response, or data on the homing of cells to the target organ, or what types of cells are involved in the rejection or tolerance, then the Mixed lymphocyte reaction is not enough.  So, it would be helpful to know what question you are asking.
  • asked a question related to Immune Response
Question
2 answers
Immunologists
Relevant answer
Answer
It would be helpful to know which reporter mice you have available, and which parasite you're studying; if this is just a general question about reporter mice or a particular set.  there are so many that without a few more details, this is a difficult question to answer.
For example, if you wanted to enumerate the T cells responding to flu, and you didn't care what kind of response they make, you could make a reporter mouse whose T cells light up when they activate CD69.  a floxed stop on the reporter color, driven by cre on a CD69 promoter would permanently color the responding cells, whereas a construct with the color on the CD69 promoter would color them temporarily.
If you wanted to know what kind of responses a particular parasite drives, then you could use reporter mice whose cells become colored when they make particular cytokines.  or you could simply look at the classes of antibodies the mouse makes (without having to use a reporter mouse). 
So it really depends on what question you're asking
  • asked a question related to Immune Response
Question
6 answers
I am studying about infections and immune response level.
Relevant answer
Answer
There will also be a lot of TNF produced!
  • asked a question related to Immune Response
Question
2 answers
My name is Jeremy, I am a senior staff nurse working in a general medical and surgical ICU in the UK.
With recently reading some work on Immunomodulatory Effect of Continuous Venovenous Hemofiltration during Sepsis: Preliminary Data.
As such I would like to ask for the readers help and advice.
With some of our septic patients experiencing acute renal failure needing CVVHDF, I have posed the question, that as yet I can find no answer to.
With 'normal' patients in ARF on CVVHDF, they would become hypothermic to a temperature of 35 Degrees or below. Hence needing active warming through a Bear Hugger and filter heater aids.
With septic patients on CVVHDF, I have seen temperatures of 36-37 Degrees with no heater elements to aid this.
If this is the case, should we not then be complementing them with heater elements to achieve an active temperature of 38-38.5 Degrees? As you may see in the normal immune response to increase an individuals temperature.
Patients are not actively given antipyretic's due to the researched based evidence of increased mortality.
If we are placing septic patients on CVVHDF, then we are actively cooling them! As such, are we not increasing the patients chances of mortality and morbidity?
Relevant answer
Answer
Fabulous question. I thought one of the reasons we avoid administering pharmocological antipyretics was their anti-inflammatory effects not necessarily the effect of reducing core temperature. CVVHDF cools by convection so the effect of cooling is confined to the reduction in temperature rather than the effect on of reduced cytokine release on the inflammatory response. 
I will continue to follow this question with interest. 
  • asked a question related to Immune Response
Question
3 answers
I am currently trying to establish a Ca2+ signaling in vivo and in vitro for T cells and also in the next step macrophages. To check the imaging properties I used a simple PMA/Ionimcyin stimulation (50ng/ml // 500ng/ml) of total T cells isolated from spleen and LN in vitro after loading the cells with OG488 BAPTA-2 for 30' at 37°C. Currently I see an astonishing demonstration of.....nothing. Could anybody suggest how to improve the method? Help will be greatly aprreciated. Best, Felix
Relevant answer
Answer
You may need to do some optimization as suggested above. Titrate the PMA/ionomycin and also do a time course. Calcium flux should happen quickly- you may be missing the peak by waiting 30 minutes to run the samples. We look within a minute.
  • asked a question related to Immune Response
Question
7 answers
Hi Everyone! We are trying to test mast cell degranulation in RBL2H3 cell line by measuring beta hexoaminidase in the supernatant. We are unable to detect the enzyme in the supernatant nor in the cell lysate. Cells are sensitized with IgE (mice anti-DNP) and with antigen DNP-BSA promoting degranulation. We didn't get any detectable enzyme in the sup so we tested cell lysate but got nothing (from the staining, granules are present). Antibody and DNP-BSA are from Sigma. Substrate
(p-Nitrophenyl-N-acetyl-β-D-glucosaminide) for the enzyme is from Millipore. We stained the cells and observed the granules after the IgE and Antigen treatments and we are not seeing degranulation. Anyone with advice? Thank you!
Relevant answer
Answer
Hi, it could be several things-buffer pHs (most likely), unhealthy cells, IgE or antigen not good / wrong concentration, lysis detergent wrong concentration. We use Tyrode's buffer pH 7.4, substrate=4.5, and stop solution=10.7. Even if you made the buffers at the correct concentration, the Tyrode's buffer pH could change over time, even with a buffering agent, so I recommend checking the pH before every experiment. Generally 1ug/mL IgE and 100ng/mL DNP-BSA are good, although the valency (number of DNPs per BSA molecule) could change these figures. 1% v/v Triton-X is good for lysis. Generally 1 hour stimulation is good and consistent. You may see some morphological changes, but don't expect to see complete degranulation using IgE and antigen. If you're using SPE-7 IgE, be careful as it can activate cells without antigen.
  • asked a question related to Immune Response
Question
2 answers
Hi, everyone,
I need to analyze miRNA expression in THP1 cells infected with Leishmania. As any experiment, I need to have a negative control of this expression. Once the THP1 will phagocyte the parasites, I would like to use latex beads in my Leishmania-free THP1 cultures, so I could create in the negative control a more similar situation to what I want to analyze, as the cells would be phagocyting something that, theoretically, won't trigger an immune response as the parasites would do. So, has anyone here used this stategy? If so, could you please share you experiences with me?
Thanks a lot.
Relevant answer
Answer
Hi, Sharon,
Yes, they are differentiated into macrophages with PMA before infection. I do not think that using inactivated/dead parasites is the best shot in this kind of analysis, because even in these conditions, the antigens are still there, and they can trigger an immune response and probably will alter miRNA expression. Thus, it would not be a negative control anymore.
  • asked a question related to Immune Response
Question
1 answer
Hi,
I want to stimulate immune response, using the antigen that combine GM1 ganglioside receptor at epithelial cells in intestine. what antigen could be used?
Thank you very much!
Relevant answer
Answer
Try Cholera Toxin B subunit. There is a lot of literature on this especially from the Czerkinsky  and Holmgren Groups
  • asked a question related to Immune Response
Question
7 answers
One of the reasons why adenovirus has short term expression is due to the strong immune response that it generates.  This anti-vector immunity usually limits the expression to ~1-2 weeks.  It is my understanding that the short duration of expression is due to CTL mediated destruction of the transduced cells.  Is this correct? I have read some papers stating that Ad can be used for short term expression of transgenes, but not mentioning that the reason for the short term expression is destruction of expressing cells.  
Short term expression is one thing, short term expression because the expressing cells are dead is another.  Can anyone shed any light on this?  
Thanks,
John
Relevant answer
Answer
Hi John,
we compared AAV and Adenovirus-5 (2nd gen.) transduction in the lung of mice and found that Adenovirus not only induced classical inflammatory cytokines (IL1, IL6, TNFa...) but also upregulated MyD88 and TLR signaling components. We also found a significant increase in CD8+ T-cells in the lung lavage samples of AdV-treated animals, suggesting that indeed cytotoxic T-cells might be responsible for the loss of AdV-mediated transgene expression. We haven't looked into direct effects of the T-cells on AdV-transduced cells, though... 
The paper is entitled "Modeling Pulmonary Disease Pathways Using Recombinant Adeno-Associated Virus 6.2."
Best wishes,
Ben  
  • asked a question related to Immune Response
Question
7 answers
We all know allogeneic immune response is one of the most powerful displays of immune reactivity. However, multiple cell types are usually involved. Has anyone ever tried or knows from literature what would happen, if allogeneic interaction ocurred solely between APCs such as monocytes??
Relevant answer
Answer
Hi Urban,
I'm not aware of the literature. I once put monocyte-derived macrophages from two different donors together in a well. Not much happened. Eventually they form clusters but syngeneic macrophages do that too after a while. You would expect allogeneic ones to start eating eachother but it wasn't that obvious in the well.. 
Try it out I would say!
  • asked a question related to Immune Response
Question
9 answers
The size of the particles and their adjuvant activity in vaccines have been controversial because some researchers showed that larger particles were more potent than smaller particles while others showed the exact opposite. There were also data showing that the size of particles did not affect the resultant immune responses and data from some studies suggested that there may be an ideal particle size with the most potent vaccine adjuvant activity
Can anybody explain about this?
Relevant answer
Answer
Hi Munir;
Particles with a size range of 10–200 nm enter the initial lymphatic vessels from the interstitial space by directly diffusing through lymphatic endothelial cell junctions. Particles of this size can also enter the lymphatic vessels after being endocytosed by dendritic cells (DCs), which then enter the lumen of the vessel by intravasation. Particles that are larger than 200 nm diffuse from the interstitial space into the lumen less efficiently with increasing size. Particles that are 500 nm–1 μm are too large to pass through endothelial cell junctions and are transported into the lymph vessel following uptake by DCs. 
Different nanoparticles have different actions on the immune cell activation, it depends on their chemical constitution.
  • asked a question related to Immune Response
Question
4 answers
We want to analyze the immune response against a protein. Our approach is to make intra-muscular injections of a plasmid that encodes for the expression of the protein. (We do not have enough AAV to do this experiment and we do not have electric devices for proceeding to an additional electric choc after injection.)
Relevant answer
Answer
You can use co-polymers ( see ref: Chèvre R, Le Bihan O, Beilvert F, Chatin B, Barteau B, Mével M, et al.
Amphiphilic block copolymers enhance the cellular uptake of DNA molecules
through a facilitated plasma membrane transport. Nucleic Acids Res 2011;39:
1610–22.
 Beilvert F, Tissot A, Langelot M, Mével M, Chatin B, Lair D, et al. DNA/amphiphilic
block copolymer nanospheres reduce asthmatic response in a mouse model of
allergic asthma. Hum Gene Ther 2012;23:597–608.)
  • asked a question related to Immune Response
Question
6 answers
Would 2-fold be a good correlation compared to MIC 24h?
Relevant answer
Answer
No, usually not. In general, the MIC is defined as "the lowest concentration (...) that will inhibit the visible growth of a microorganism after overnight incubation" (see link).
Be aware, that MIC determination is a highly standardized procedure, and that labs doing MIC determination are advised to adhere to the respective guidelines. CLSI in its M07-A9 document gives an incubation time of 16 to 20 hrs, except for some fastidious organisms or difficult-to-detect resistance mechanism. Specifically, for Staph, the MIC should be read after 16 - 20 hrs (except Vanco and Oxacillin, which require 24 hrs of incubation), as recommended in the CLSI M100 guideline.
If you incubate longer, you are running into problems with growth of the fraction of bugs, which was not initially inhibited and of those bacteria which regrow after post-antibiotic effects fade out.
  • asked a question related to Immune Response
Question
2 answers
We are studying mature T cell proliferation and migration in vitro and we are trying to find an infection mouse model in which we could compare the immune response when either the proliferation or the migration of T cell are impaired. Does anyone have any suggestion about which infection we could use? Do you think that a systemic infection, like salmonellosis, could be useful to analyse a possible defect in T cell proliferation upon activation? On the other side, could a local infection like pulmonary influenza be the best to analyse T cell infiltrations? Better suggestions? Thanks for help.
Relevant answer
Answer
You may try an autoimmune disease model, pMOG35-55-induced experimental autoimmune encephalomyelitis (EAE) on B6 background.  In this model, pMOG-35-55-specific T cells are first activated and expanded upon immunization, and then migrate to CNS to induce inflammatory response in CNS and cause overt clinical symptom.  If the genetic-modified mouse strain has T-cell defect (either proliferation or migration, or both), it should be resistant to EAE induction (the WT control mice should develop EAE).
  • asked a question related to Immune Response
Question
6 answers
Been doing beta-hexosaminidase release assay but the results hasn't been promising. Firstly, there's no colour development in the supernatant and secondly the release has been below 10% for all groups. 
Tried doing ELISA but there wasn't any difference between the normal and induced groups. 
What went wrong - the cells, the antibody, buffer??
Relevant answer
Answer
I've done beta-hex assays with RBLs for years. There are many potential problems, but pH is the most likely. RBLs are unresponsive at low pH. If you're using Tyrode's buffer the pH should be close to 7.4. Even with HEPES I've noticed that the buffer pH will decrease over time if the buffer isn't frozen. I used to check the Tyrode's buffer pH every time I did the assay. I've never seen the substrate (pH 4.5) or stop solution (pH 10.7) go off pH. This could explain your ELISA result as well if you used the same buffer. What cytokine did you assay for?
  • asked a question related to Immune Response
Question
8 answers
I would like to check if my PGLA injections are influencing the mice immune response. I collected sera at some time points. For you, what is the method of choice? And which cytokines do I have to measure?
Thanks for sharing your expertise.
Relevant answer
Answer
Hi Sarah ,
You can assay pro-inflammatory cytokines in the injection site via assay on tissue homogenates and also you can monitor inflammatory cell trafficking in this micro- environment via pathological smear. 
Regards
  • asked a question related to Immune Response
Question
1 answer
I know MCSFR (CD115 or CSF1R) mRNA is expressed by GMP (granulocyte-macrophage progenitor) and CMP (common-myeloid progenitor), but what about protein? Is CD115 detectable on the cell surface of GMP and CMP?
Relevant answer
Answer
This work might help!
www.ncbi.nlm.nih.gov › NCBI › Literature › PubMed Central (PMC)
by YK Lieu - ‎2012 
Impaired adult myeloid progenitor CMP and GMP cell function in conditional .... No abnormality was detected in the pIpC-treated mybf/+/MxCre mice (data not shown). ... Cell surface expression of CD11b, CD41 (a marker of megakaryocytic ... in the CD11b+Gr-1- compartment also expressed elevated surface CD115 and ...
  • asked a question related to Immune Response
Question
3 answers
When the animal is challenged intranasaly or intratracheally will there be any clinical signs in animal? I challenged calves intranasally with 5ml of inoculum containing 109 cfu/ml but observed no clinical signs. Also no clinical signs were produced when the calves were challenged intratracheally with 109 cfu. So in such case will there be any immune response?
Relevant answer
Answer
I agree with dr Atef answer. It is important to exclude the presence of Pasteurella before starting experimental infection experiment. You may observe some respiratory signs if the animals are free of Pasteurella. If they are natural carrier of some strains and you use a laboratory strain  immune response  could be unsignifiant, since natural strains of Pasteurella may inhibit growing of laboratory strains
  • asked a question related to Immune Response
Question
2 answers
I would like to know, the relationship between the G-CSF and TH17 in cancer. Wich one influence the other, and how they act in anti-tumoral immune response. Some recent articles will be useful. 
Thanks.  
Relevant answer
Answer
Hi   see this paper may help your work
  • asked a question related to Immune Response
Question
10 answers
To dampen immune responses, the anti-inflammatory cytokine TGF-beta can be produced by macrophages. However few good detection methods are present or reproducible to detect this cytokine. Anyone used a good antibody to detect the secretion of this cytokine (or a publication)?
Relevant answer
Answer
I suggest also R&D
  • asked a question related to Immune Response
Question
8 answers
In several clinical settings such as pancreatitis and trauma, a secondary infection can be frequent and often causes death. I wonder if the body in a hyperactivated status would amplify the detrimental effect of bacterial infection. Is there any literatures on this aspect?
Relevant answer