Immunology - Science topic

Anti-HBcAb (antibodies against hepatitis B core antigen) is produced by the immune system in response to exposure to the hepatitis B virus (HBV). When HBV infects hepatocytes (liver cells), the cells produce hepatitis B core antigen (HBcAg) as part of the viral replication process. HBcAg is not released into the blood in significant quantities; instead, it remains within the infected hepatocytes or is present in viral particles.The presence of HBcAg within infected hepatocytes triggers an immune response in the body. B cells, a type of white blood cell, recognize HBcAg as foreign and start to produce antibodies against it. These antibodies, referred to as anti-HBc antibodies, are then released into the bloodstream. The presence of anti-HBc antibodies can be detected through blood tests and indicates exposure to the hepatitis B virus, either currently or in the past.

Autoimmune disorders can significantly impact fertility, affecting both males and females by interfering with reproductive functions. These conditions can lead to infertility through various mechanisms, including hormonal imbalances, thyroid autoimmunity, and direct effects on reproductive tissues. Studies highlight the importance of identifying and managing autoimmune disorders in individuals facing infertility, suggesting that tailored therapies, including cell and gene therapies, might improve fertility outcomes in affected individuals.

Optimizing the purification and differentiation of dendritic cells (DCs) in vitro is crucial for obtaining a homogeneous and functional population for research or therapeutic purposes. Here's a step-by-step guide to optimizing this process:

1. Source Material Selection: Choose PBMCs, bone marrow, or umbilical cord blood based on availability and experimental needs.

2. Isolation of Precursor Cells: Use techniques like density gradient centrifugation or magnetic bead separation with sterile protocols.

3. Differentiation Protocol Optimization: Cultivate precursor cells with specific cytokines and growth factors, adjusting concentrations and timing for optimal differentiation. Monitor differentiation using flow cytometry.

4. Maturation Induction: Stimulate immature DCs with appropriate stimuli like Toll-like receptor agonists or cytokines, optimizing concentrations and duration.

5. Culture Conditions Optimization: Maintain DC cultures in optimized media with proper supplements, adjusting temperature, pH, oxygen levels, and cell density as needed. Consider using feeder cells or specialized systems.

6. Quality Control and Characterization: Perform thorough phenotypic and functional characterization using flow cytometry and functional assays to ensure purity, phenotype, and functionality of generated DCs.

7. Cryopreservation Optimization: Develop optimized cryopreservation protocols for long-term storage, ensuring viability and functionality post-thawing.

The Sterile filtration of antibodies is to be injected intraperitoneally (IP) into mice for neutralization studies to ensure the absence of microbial contaminants and to reduce the risk of introducing infection or other complications into mice. However, whether this is necessary depends on various factors including the source and handling of the antibody, the experimental setup, and the specific requirements of the study.

Granulocytes, such as neutrophils, eosinophils, and basophils, are a type of white blood cell that plays a key role in the immune response, particularly in combating infections. Activated granulocytes undergo various changes in morphology, surface marker expression, and functional activities. Here are several methods to assess the activation of granulocytes from peripheral blood:

1. Morphological Assessment

2. Flow Cytometry

3. Measurement of Reactive Oxygen Species (ROS) Production

4. Assessment of Degranulation

5. Functional Assays

6. Quantification of Cytokine Secretion

7. Gene Expression Analysis

Yes, B and T cells can be raised against pathogens that may not have been initially detected by the innate immune system. The adaptive immune system, comprising B and T cells, is adept at recognizing specific antigens presented by pathogens, even if they initially evade detection by the innate immune system. Once activated, B cells differentiate into plasma cells, producing antibodies targeting the pathogen, while T cells undergo clonal expansion and differentiate into various effector cell types. This adaptive immune response, along with the generation of memory B and T cells, contributes to long-lasting immunity against pathogens, irrespective of their initial evasion of the innate immune system.

Dear Esteemed Colleague,

Greetings. I trust this message finds you deeply engaged in your research and seeking answers to complex questions within the realm of genetics and molecular pathology. Your inquiry regarding the potential role of infection in causing desmin mutations in myofibrillar myopathy is both intriguing and indicative of a keen scientific mind exploring the multifaceted nature of genetic disorders.

To address your question with the precision and clarity it deserves, it is crucial to first understand the nature of myofibrillar myopathies and the role of desmin within this context. Myofibrillar myopathies are a group of neuromuscular disorders characterized by the progressive weakening of muscles and the disintegration of muscle fibers at a cellular level. Desmin, a type of intermediate filament protein, plays a pivotal role in maintaining the structural integrity and function of muscle cells. Mutations in the DES gene, which encodes the desmin protein, are directly linked to certain forms of myofibrillar myopathy.

The genesis of these mutations, particularly those affecting the desmin protein, is primarily genetic, resulting from inherited or de novo mutations in the DES gene. These mutations lead to the production of an abnormal desmin protein, which disrupts the normal architecture of muscle cells, leading to the symptoms associated with myofibrillar myopathy.

Addressing the specific question of whether an infection could cause desmin mutations, it is essential to differentiate between the origins of genetic mutations and factors that may exacerbate the phenotype of a genetic disorder. Genetic mutations, including those affecting the desmin gene, arise from alterations in the DNA sequence. These alterations can be inherited from parents, occur spontaneously during DNA replication, or be induced by certain environmental factors, such as exposure to specific chemicals or radiation. Infections, while capable of causing a wide array of health issues, do not directly induce genetic mutations in the DNA sequence of the genes like DES. However, it is conceivable that certain infections could exacerbate the clinical manifestations of myofibrillar myopathy in individuals already predisposed or carrying a desmin mutation, by stressing the muscular system or triggering inflammatory responses that may further compromise muscle function.

In conclusion, while infections can have significant impacts on overall health and may interact in complex ways with genetic disorders, the mutations in the DES gene that cause myofibrillar myopathy are not directly caused by infections. The mutations are genetic in origin, and the relationship between infections and the severity or progression of myofibrillar myopathy would be more accurately viewed through the lens of infection exacerbating pre-existing conditions rather than causing the genetic mutation itself.

I hope this elucidation addresses your inquiry comprehensively. Should you have further questions or require additional clarification, please feel free to reach out.

Warm regards.

This protocol list might provide further insights to address this issue.

Thank you very much :)

can you kindly show the graph? That will aid understanding of the question.

You might try an anti-SRY antibody as for sperm cell sorting; but honestly, I would rather flow sort the various cell populations and do FISH with a Y-chromosome-specific probe.

Hi Annam. Greco

just on a side note. You can run the Olink assays locally either at the

Immunology and Vaccinology laboratory at the Bambino Gesù Children's Hospital, University of Rome Tor Vergata or IRCCS Centro Cardiologico Monzino.

All the best & kind regards,

Michael

Normally, to get a good immune response, you need both T-cell and B-cell epitopes in your Immunogen. You may get a T-cell independent immune response if your immunogen is highly polymeric, but in this case the response will not progress beyond IgM. By coupling a hapten to a suitable carrier protein you both provide T-cell epitopes and multimerization of the hapten. The response will usually consist of a mixture of B-cells/antibodies with different specificity: some only recognizing the hapten in the context of the peptide sequence it is coupled to, others may also recognize the free hapten. Since you randomly couple the hapten to different sites of the carrier, those that recognize the hapten independent of the sequence context have an advantage, since they "see" a larger antigen concentration. On the other hand, since the binding affinity of an antibody is limited by the contact area, antibodies recognizing more than just the hapten may achieve higher affinities. In consequence, the antiserum generated will most likely contain both antibodies against the hapten alone and antibodies only recognizing the hapten in the context of the carrier protein sequence it was coupled to. The proportion of the two types of antibodies in the serum may vary between different animals injected with the same immunogen as well as in the course of the immune response. If you generate monoclonal antibodies, you can screen for clones that fit your requirement.

Desminopathy, also known as desmin-related myopathy (DRM), is a rare genetic muscle disorder that affects the protein desmin. Desmin is an essential component of the intermediate filaments that provide structural support within muscle cells. Mutations in the DES gene, which codes for desmin, lead to disruptions in the normal structure and function of muscle fibers.

The muscles you mentioned - Semitendinosus, Gracilis, and Sartorius - are often affected at the onset of desminopathy due to their specific characteristics and biomechanical roles.

1. Semitendinosus: The semitendinosus is one of the three hamstring muscles located in the back of the thigh. It plays a key role in knee flexion and hip extension. The semitendinosus muscle is frequently involved in desminopathy due to its high proportion of slow-twitch muscle fibers, which are more vulnerable to desmin-related abnormalities.

2. Gracilis: The gracilis muscle is a long, thin muscle located in the inner thigh region. It is involved in hip adduction and knee flexion. Similar to the semitendinosus, the gracilis muscle also consists of a high proportion of slow-twitch muscle fibers, making it susceptible to desmin-related abnormalities.

3. Sartorius: The sartorius muscle is a long, strap-like muscle that runs diagonally across the front of the thigh. It plays a role in hip and knee flexion and also assists in thigh rotation. The sartorius muscle is affected in desminopathy due to its similar composition of slow-twitch muscle fibers.

The predilection for these specific muscles in desminopathy may be attributed to their fiber type composition and the mechanical stress they experience during certain movements. However, it is important to note that desminopathy can affect other muscles as well, and the degree and pattern of muscle involvement may vary among individuals with the same genetic mutation.

It is advised to consult with a medical professional or genetics specialist for a more accurate assessment of muscle involvement and management of desminopathy.

Dear Dr. Suha Abadullah Alfakhar

The reason for this increase in TNF-alpha levels is not fully understood, and the answer may vary between individuals. It has been suggested that inhibition of TNF-alpha by biological therapies may disrupt normal regulatory feedback loops, leading to an unintended increase in TNF-alpha production. It is important to note that the clinical significance of increased TNF-alpha concentrations during anti-TNF therapy is not completely clear and does not necessarily imply treatment failure. Monitoring patients for both clinical symptoms and laboratory parameters is important, and adjustments to the treatment plan can be made based on individual patient response.

AB Bayazid

Thanks!

Dear Sina Tadayon

I briefly wanted to to reach out concerning your inquiry. I agree that your Lin‐cKit+Sca1+CD150+CD48- cells are the LT-HSCs whereas the Lin‐cKit+Sca1+CD150+CD48+ constitute a hematopoietic progenitor population which will differ in their self renewal capacity when transplanted in competitive bone marrow transplantation experiments. For details please see Figure 2 and supplementary data in:

In addition, please bear in mind that these populations will differ between immunocompetent and deficient mouse strains, underlying their hematopoietic defects. For details please consider recent work from the Levesque lab.

I hope that helps.

All the best & good luck with your experiments,

Michael

The fist thing to know the vaccine strain used dose ,titer ,effect of booster dose,then study immunological status of vaccinated person before vaccind and after vaccination,need for booster dose or not the select your aim

At this time ( Sept,2023) Dengue crisis is appearing worsen in Dhaka , Capital of Bangladesh. Patients are being treated by paracetamol only but high fever not responding, rather complications are encouragingly increasing. I believe, complications are due to cytokines or other chemical mediators released immunologicaly, attached to the target cells or tissue. I think, the half life of these chemical mediators is short and as soon as steroid starts acting against inflammation, the temperature rapidly subsided. As viremia in dengue may persist for several days steroid should be continued till the chance of inflammation. Rapid remission of temperature with a single dose of prednisolone within 6 to 8 hours is a miracle, though cause of this action is a matter of much investigation. However the information should be rapidly disseminated among the medical professionals for the benefit of the vulnerable patients.

Some immunologists believe that, after each round of antigenic stimulation, the progeny of memory B cells are more likely to become plasma cells (which die) than new memory cells (which survive). In vivo, this could translate into an increased number of effector cells in the secondary and subsequent responses, and a control on the possible overexpansion of one particular memory B cell clone. However, it also means that the host might one day no longer have memory cells of this clone to call upon when the relevant pathogen strikes. In this “decreasing potential hypothesis,” immunological memory is ultimately limited. In addition, for reasons that are not yet understood, memory cells specific for different antigens have different life spans. These variations have implications for how frequently a booster shot must be given to ensure complete vaccination against a particular pathogen.

Memory cells are incredibly powerful tools for our immune system and can be very long-lived, with studies showing memory B cells for smallpox persisting at least 60 years after vaccination and for Spanish flu at least 90 years after the 1918 pandemic.

Memory B cells have several unique features including long lifespan, high sensitivity to low doses of antigen, quick and robust proliferation, and rapid differentiation into plasma cells that produce high-affinity antibodies during the secondary response.

In this sense, Md Mominul Islam , memory cells can be understood as ‚intelligent‘ or ‚learning‘ cells, imo.

Greetings! I'm delighted to share my insights and experiences with you in the realm of transcriptomics, immune responses, and machine learning. When comparing the performance of random forests and decision trees in predicting immune responses using transcriptome data, I've found the following performance metrics to be most informative:

When working with complex gene expression data, it's essential to consider the biological relevance of the performance metrics. For instance, in some cases, a high accuracy might not necessarily translate into biologically meaningful results. Instead, focus on identifying models that capture the underlying biology effectively, such as those that distinguish between different immune cell subtypes or predict functional pathways.

To optimize model parameters and prevent overfitting, follow these best practices:

By considering these performance metrics, biological relevance, and optimization strategies, you'll be well on your way to developing robust and reliable machine learning models for predicting immune responses using transcriptome data.

There are some differences between visual inspection guidelines for sterile immunological veterinary products compared to human products:

So in summary, while the general principles of visual inspection for defects and particulate matter apply to both human and veterinary sterile immunologicals, veterinary products lack standardized global guidance. Acceptance criteria may be more flexible based on a case-by-case risk assessment.

I hope this helps

Hi Sam Marcs, the issue you encountered with counting the magplex beads using a hemocytometer can be challenging without a dedicated cell counter. However, there are a few potential reasons for the discrepancy you observed:

Also, considering that your luminex test results were consistent with the expected concentrations, it suggests that the bead preparation and dilution were likely accurate. However, it's always a good practice to optimize your counting method and validate it against alternative techniques if possible.

If access to a cell counter is not available, you can explore other methods to estimate bead concentration, such as using a spectrophotometer to measure the absorbance at specific wavelengths or employing imaging software for automated counting based on images of diluted bead samples.

Cheers!

-H-

Here are some research topics in the field of immunology that do not involve wet lab work:

Dear Abdulla,

I have found several articles that are related to the topic you are interested in. Here they are for your reference. Enjoy your reading!

1. Emerging Immune Biomarkers for Prognosing the Outcome of COVID-19 Patients

2. Comparative Immunological Analysis of Vaccinated Versus Recovered COVID-19 Patients

3. Molecular Comparison of Recovered and Vaccinated COVID-19 Patients

4. Transcriptomic Profiling of Recovered and Vaccinated COVID-19 Patients: Insights from a Comparative Analysis

5. Comparison of Immune-Related Genes of Recovered and Vaccinated COVID-19 Patients

6. Altered Innate Immune and Inflammatory Responses in Response to Vaccine Versus Natural Infection of COVID-19

7. Serological and Molecular Evaluation of Vaccinated and Recovered SARS-CoV-2 Patients

8. A Comprehensive Comparison of Vaccinated versus Recovered COVID-19 Patients

9. Comparative Immunological Evaluation of Vaccinated and Naturally Recovered COVID-19 Patients

10. Differential Analysis of Immuno-molecular Parameters Between Vaccinated and Naturally Recovered SARS-CoV-2 Patients

Best regards, Saif

Spectratyping is indeed a commonly used method for TCR repertoire analysis, particularly for assessing the clonality and diversity of T-cell populations. However, it is important to note that spectratyping primarily provides qualitative information about the distribution of TCR lengths and does not offer detailed sequence-level information.

Several software tools are available for TCR repertoire analysis, which can be used in conjunction with these sequencing methods. Here are a few popular ones:

Top of Form

Dear professor,

sorry but I have many problems to partecipate at the Conference because of my cronic heath problems.

All my best, Catina Feresin

cortisol, testosterona , Iga salivar, glutationa reduzida (GSH), glutationa peroxidase (GPx), catalase (CAT), proteína carbonilada (PC), substâncias reagentes ao ácido tiobarbitúrico (TBARS), atividade da enzima mieloperoxidase (MPO), metabólitos do óxido nítrico (NOx), proteína C reativa (PCR), spo², il-1b, nfkb, tnf-a

There are a number of factors to consider and certainly there are ways to preserve and store plates for longer periods of time. However, in your case the key question is the stability of the antigen. If you know the antigen is stable, then you should be fine to store for several days in the coating buffer before proceeding with your ELISA. I have done this with several antigens and have never observed an issue with using the plate within a week. Hope this helps.

Hi Keshav Bhatnagar ,

This might not be exactly what you need, but should give you a reasonably good idea what approach to take based on the equipment used in the protocol you are trying to replicate and the equipment available to you:

http://vesicles.niifhm.ru/

Some of the theory to help you understand what calculations are being done in the background can be found here:

https://handling-solutions.eppendorf.com/sample-handling/centrifugation/this-and-that/detailview/news/adapting-centrifugation-time/

Hope this helps!

The shift between two opposite direction of change is the rule for regulatory genes that work as 'toggle.switches' in which the biphasic alternation of two conditions is the basis for a sort of digital control of biological regulation. It is not by chance that a great part of toggle-switches are retroviral origin sequences that mirror the lytic-lysogenic phases of viruses and phagi, see:

If you are talking about using the antibodies as the affinity matrix (such as in immunoaffinity chromatography), I would suggest that the way to avoid damage to the antibodies from proteases in a crude extract is to save the immunoaffinity chromatography for the last step in the purification, at which point the protease activity from the crude extract should be greatly reduced. Begin the purification with ion exchange, hydrophobic interaction, and/or gel filtration chromatographies. Use a protease inhibitor cocktail when making the crude extract.

"Genetically restricted" in the context of immunology refers to the idea that an individual's immune system is only able to recognize and respond to certain types of pathogens, or disease-causing agents. This ability is determined by the genes that an individual inherits from their parents, which encode for the proteins that make up the immune system. These proteins, called antigens, are used by the immune system to identify and target pathogens. An individual's immune system is only able to recognize and respond to a limited range of antigens, meaning that it is genetically restricted in its ability to defend against certain types of pathogens. This is why some people may be more susceptible to certain types of infections or diseases than others.

1. Develop an Area of Expertise: Develop a specific area of expertise related to your research interests. This can be done by reading relevant books, journal articles, and other resources in your chosen field.

2. Network: Networking is important for any researcher. Reach out to colleagues, professors, and other experts in the field for advice and to stay abreast of the latest developments.

3. Attend Conferences: Attend conferences related to your research interests to stay current on the latest developments and to meet other researchers in the field.

4. Join Professional Organizations: Joining professional organizations related to your research interests can help you stay current on the latest developments, as well as provide you with access to valuable resources.

5. Develop Collaborative Relationships: Connect with other researchers for collaboration opportunities. This can help you develop new ideas, gain new perspectives, and learn from one another.

6. Utilize Online Resources: Take advantage of the wealth of information available on the Internet. Use search engines, databases, and other resources to stay up to date on the latest developments and research findings.

7. Publish Your Work: Publishing your work is a great way to make a name for yourself in the

Yes, artificial intelligence (AI) techniques can be used in immunology to analyze and interpret large amounts of data, such as genetic sequences and protein structures. AI can be used to identify patterns and correlations in this data that may not be easily visible to the human eye, and to make predictions about how the immune system will respond to different stimuli. For example, AI can be used to predict the likelihood of an individual developing a specific autoimmune disorder or to identify potential targets for immune-based therapies. AI can also be used to analyze the results of immunological experiments and to develop new computational models for studying the immune system.

As far as I know all lymph nodes are almost same other than sizes !

From our experience, the concentration data of the manufacturers are oversized. We use only 1 μl on about 1x10^6 cells in 100 μl. So far, we have always been able to achieve good FACS results, whether with antibodies for mice or humans. Decisive is of course the amount of your marker to be detected and the correct selection of fluorochrome which have a different intensity. Therefore, a prior establishment of the fluorochrome composition is always advisable. Regards

Dear Dr. Edmund Pool

That's great and much appreciatable.

Hello Mahmoud Gharieb

It will depend on the concentration-exposure relationships that can differ between individuals, due to acquired or genetic host factors. The type and intensity of interaction between the drug and target may relate to both the dose and duration of treatment.

Some off-target effects are both directly immune-mediated and associated with immunological memory of varied duration (drug hypersensitivity), whereas others without immunological memory might have an immunological phenotype, such as non-IgE-mediated mast-cell activation seen with the use of certain antibiotics like fluoroquinolones.

Best.

Dear Jean-Jacques Pin Jacques, I have a question related to the supplement. Is it okay for using BM-Condimed H1 for human hybridoma production?

Sorry for asking through this discussion

Hello Hyungjoo Kim

Yes, I do have some ideas that may be of interest to you.

As you may be knowing that T-cell therapy trials have been successful in certain blood cancers. But what about cancers that are solid tumors?

Solid tumors pose challenges for immunotherapies that aren’t factors in liquid tumors, where the cancer cells are in the bloodstream and are more accessible to immune cells.

There are a lot many factors within the tumor microenvironment that can shut down T cells, and these factors may/may not be the same for every solid tumor. There might be certain types of immune cells which might hinder this process, or cancer cells themselves might put up a fight by sending immunosuppressive signals that is likely to further dampen the anti-cancer immune responses.

What are your thoughts on this?

Another interesting idea! Still unanswered.

Can cancer be prevented by training our own immune system?

For this purpose, we would have to characterize the genome and the immune environment which is associated with precancerous growth. Find potential targets and bombard them with immune boosters or develop vaccines to prevent cancerous growth.

Best.

Hi Tulika, I am also switching my studies to 384 well plates. I will be using suspension cells not adherent. I am wondering what was your experience with 384-W plate.

Thanks a lot in advance,

detection the low concentration (as you said) of anti-dsDNA IgE depends on the sensitivity of your ELISA kit. but you also can make a pilot test by using different dilutions to find out the best dilution concentration that you should use it.

Hi Paulius Valiukevicius

I would suggest you pass your cells through a 40 um cell strainer prior to your FACS analysis. That should help with your clumping issues, but you might loose some cells.

Please note that human T cells express human leukocyte antigen (HLA)-DR-alpha (DRA) upon mitogenic or antigenic stimulation, which makes them incredibly sticky, especially when you culture bulk PBMCs without separating the different cell types prior to setting up your cultures. I hope that helps.

All the best & good luck with your experiment.

Michael

Hi Sudeep Kumar Maurya

I commonly use CD3 to gate T cells; then CD4 and CD8 to gate T helper cells and T cytotoxic cells, respectively; Then using a CD3+CD4+ gating (Th cells) you could use CD25 and CD127 in combination to distinguish between effector T cells (CD127+CD25low) and regulatory T cells (CD127low CD25high).

You can check one of my articles for that: doi: 10.1038/s41598-019-43622-8

Best

Juanjo

Hi,

Check the references below:

HCMV UL83/pp65 (NLVPMVATV)

Reference: UNIPROT https://www.uniprot.org/uniprotkb/P04439/entry

CMV pp65 peptide NLVPMVATV (HLA-A*0201) is a single peptide for stimulation of T cells. The peptide from human cytomegalovirus (CMV) phosphoprotein (pp65) is synthesized as it is presented by the MHC class I HLA-A*0201 allele and can be used for targeted in vitro generation and expansion of antigen-specific CD8+ T cells.

Reference:

"Although the anti-pp65 antibodies and the pp65 antigens are detected in immune-depressed patients with active viral infection [16], the antibody response to the pp65 antigen in normal infected individuals is not always detectable by immunoblotting [17]. It has been reported that the pp65 antigen is targeted by a cell-mediated immune response and that its vaccination can induce a pp65-specific CTL response"

Reference:

or https://academic.oup.com/cei/article/143/1/167/6440089

Hope it helps,

Best regards

Tomasz

No, there are essentially 3 major types of immune response to infection: an antiviral response which illicits interferon, a pro inflammatory response that makes il1, il6, tnfalpha etc wich is normally directed against bacterial and fungal infections ( and can lead to cytokine storm) and then the allergic response directed against allergens like ova that generates il4, il10, il13.

In mouse models, balb/c mice are typically used to study allergic responses as thy are inherently more allergic and c57bl/6 are used to study proinflammatory responses and diseases because they are inherently more prone to this type of immune reaction.

It's better if you provide more details about your questions. based on my experience, the flowjo software can show distinct populations based on their marker expression intensity. in a flowjo plot, one dot represent one cell, so you can also get the percentage of every population you have. these are basic functions that flowjo have, for more detailed, you can search on the official FLOWJO website or there are some useful youtube video that show how to analyse flow data by flowjo.

Hi, also you could give a try to ENCODE. Check out the following link, there is an overview of the immune cell lines profiled by different omics.

Not really. There's been a ton of effort going into developing tools to do this but most are in early stage development and not ready for routine usage. The main issue is a lack of training data that can be used to refine these algorithms.

There are new in vitro high-throughput functional TCR antigen profiling techniques that have come online in previous years, including the one we developed: https://www.researchgate.net/publication/336310777_Rapid_selection_and_identification_of_functional_CD8_T_cell_epitopes_from_large_peptide-coding_libraries

These approaches are powerful because they let you directly test large libraries of candidate sequences in an unbiased way. If you have any interest in trying out our method, send me a DM and we can talk further.

1. Equip and Encourage

As leaders, we should make time to debrief college students about their experiences. Talk to them through their missions experience, and while doing so, inspire them to follow what they have discovered back on campus. Provide avenues for scholar missionaries to use what they have realized in their mission experience on campus and thru your ministry. Intentionally suppose outside the field about how to connect their special journey to special ministry opportunities.

Lauren and Ryan began dating before Lauren left for a semester of missions journey in South Asia. While she used to be serving in India, Ryan looked around to see the multitudes of Indian students on campus. He started apartment ministries with these students which eventually supplied avenues for Lauren to make investments in ministry to South Asians when she lowers back home.

2. Challenge

Hold college students accountable. Help them be accurate stewards by means of difficult them to use the journey God has given them. When time and college lifestyles seem to take over, mission them to stay the course.

The testimonies stated above haven’t stopped. Haley has lately back to East Asia – this time as an English teacher. She has had the probability to construct some of those relationships that started out on her campus in Alabama, through connecting with these equal college students in East Asia! While Ryan and Lauren are preparing for the subsequent steps overseas, they have maximized their professions to be planted in a predominant US town among South Asians here. They are taking this time to study more about the subculture and put it together for a probability to serve in South Asia.

Progenitor cells in the tumor are usually Granzyme K⁺ and slowly proliferate. As the cells flip towards transitory/effector-like they become Granzyme B⁺ and stay Granzyme B⁺ as they further exhaust. So in essence there are Ki-67⁺ (with the capacity to proliferate) cytolytic cells and non-proliferating/terminally exhausted subsets.

It is stochastic, or random. The receptor repertoire based on vdj, alpha beta v delta gamma and n and p additions result in a nearly unlimited. Umber of potential receptor rearrangement. The naive t cells enter secondary lymphoid organs at the instruction of chemokines and 4andomly test against antigen presented in the context of correct mhc. In this scenario, it I'd the antigen that selects the correct tcr, resulting in clone proliferation. The t cell is the passive player.

Can I also get recommendations for universities offering masters in the same subject abroad as well. I am also looking for those options as well.

Are you using this as a stain for dead cells?

7AAD is a fluorochrome used as a marker of apoptosis, and is typically added as a last step.

I don't think you need to remove before your run, however, I would consider doing a longer incubation.

Take a look at this protocol: https://www.thermofisher.com/document-connect/document-connect.html?url=https%3A%2F%2Fassets.thermofisher.com%2FTFS-Assets%2FLSG%2Fmanuals%2Fmp35123.pdf

Hi André Cronemberger Andrade I read your comment and I would like to know if you have any protocol for what you mention about sensitizing the elisa plate to later detect surface proteins in exosomes. Do you use elisa plates of any particular brand?

I would appreciate it if you could help me, thanks in advance.

Also these are good:

1- Writing Research Papers. Student's Book: From Essay to Research Paper

Dorothy E. Zemach, Daniel Broudy, Chris Valvona - 2013 - 128 pages

2- English for Writing Research Papers (English for Academic Research)

Part of: English for Academic Research (11 Books) | by Adrian Wallwork | Mar 3, 2016

3- https://www.scribbr.com/category/research-paper/

Hi Sandeep Kumar Dhanda

Just visit the website of IEDB Analysis Resources. You can see the role of Machine Learning Algorithms in reverse vaccinology, mainly epitope predictions.

Kalyanaraman

As far as I understand, your data points/conditions are nested within individuals (donors) creating non-independence. In other words, the same individuals provide data for each condition. If that is true, then a paired test such as repeated measures ANOVA would be most appropriate. One way to examine this is by looking at whether the observations across conditions are correlated. For paired observations, you will typically find a positive correlation of the DV scores across conditions reflecting the non-independence of the data points.

Hello Hasanthika C. Adhihetty

Immunology is a branch of biomedical science that deals with the study of immune system in health and disease. It has applications in various disciplines of medicine like oncology, virology, bacteriology, and organ transplant. With a Masters in immunology, you have various options open like Clinical Research Assistant in hospitals and clinics, technical support in clinical instrumentation like flow cytometry and ELISA-based techniques, and as a teaching faculty in universities.

Immunology is an interesting subject. If you wish and have a liking for immunology you can study further and complete your PhD in immunology and become an immunologist. This will be an added advantage as it will help you acquire a much higher position in academics, research or pharmaceutical industry.

All the Best.

Have a look at these useful links.

Kindly visit...

Sorry it is outside my aera of expertise

Let's change it this way, Its possible to get serum from EDTA-K₃ plasma? the answer is yes.

Briefly, Centrifuge sample for remove of blood cells and keep buffy coat plasma at -20 °C. After thawing of the sample, dispense in another tube 1.0 ml of plasma add 26 μl of 1 M CaCl₂ and incubate overnight in +4 °C. A clott is present at the end of incubation that must be squeezed and removed, obtaining clotting factors free serum.

The difficulty is that if Antibodies bind nicely to your antigens it's usually that they are very affine to them and subsequently separating them is sometimes denaturing the antibodies themselves. They are some Mild elution buffer made up by Pierce you might give a try on that.

Other than that is to use maybe a higher salt content to protect antibodies but this is rarely working perfectly

Some more litterature you might want to read

China is the best choice, there are vast opportunities for the young

Hi,

Very important question. Moreover, the question is why (.... from my current knowledge please correct if I'm wrong) from more less 100 mAbs approved by FDA only 2 are bispecific ..... I think another trend is competitive for the "multispecificity" of mAbs. Its cocktails of mAbs (cocktails therapy). See some examples below:

1 https://www.fda.gov/news-events/press-announcements/fda-approves-first-treatment-ebola-virus

2 https://www.fda.gov/news-events/press-announcements/coronavirus-covid-19-update-fda-authorizes-monoclonal-antibodies-treatment-covid-19-0

3 https://www.fda.gov/media/106517/download

4 https://www.fda.gov/drugs/drug-safety-and-availability/fda-authorizes-bamlanivimab-and-etesevimab-monoclonal-antibody-therapy-post-exposure-prophylaxis

I think that cocktails therapy will be a more dynamic trend than "multispecificity" (because of higher and higher availability of different monospecific monoclonals)... and many other reasons,

Best regards

Tomasz

Fcs alternatives for HL 60 neutrophil like cell line is FPS (slightly different).

FoxP3 + should be CD4+CD25+ Tregs. of Couse,FoxP3 should overlap with CD3+ T cells. usually CD3 should be expressed on the cell surface, while FoxP3 should be expressed in the nuclear of cells. they should co-expression in one cells. if you see they are differently localized. it means it is a unspecific binding and it is not correct. I often see some published paper they are wrong! confocal will detect them more accurately!

Overall, PRNT has been reliable and a mainstay for virology and vaccine development. Here are the limitations I've found personally:

1) The throughput of the assay is somewhat limited due to the necessity of plaque counting as opposed to an automated read-out of reporter gene activity. I've really only done manual counts, and it looks to be somewhat difficult to automate the process. There are scanners with software for counting, but the visibility of the plaques and contrast with the uninfected monolayer can be limiting and the software may not give accurate counts. Related to this is the need to have the plaque size of the target virus being uniform so that counting is accurate.

One workaround is to examine the dilution series of each antibody and count only the two wells that bracket the endpoint, such as the 50% reduction in plaque counts. That can often be done in a hurry or to get preliminary data around the endpoint titer, but you lose a lot of information that way.

Another caveat for plaque counting either by human or machine is the necessity for cell monolayers to be uniform and complete enough in each well to get accurate plaque counts. If the cells do not form a 100% uniform monolayer by the end of the plaque growth against which the plaques can be counted, then it gets tricky, especially if the plaques/monolayers are stained with a dye rather than by immunochemical staining which may be better at showing the infected cells on a bad monolayer but is more time intensive.

2) The biosafety containment needs to be commensurate with the target virus biosafety. Often, researchers use the replicating virus of interest in the PRNT as opposed to a pseudovirus or chimeric virus which can be made to have lower biosafety requirements but will often have a reporter transgene if one is going to all the trouble of constructing these.

3) For some viruses that are more difficult to neutralize than others, it can be a little tricky to normalize the input virus between assays. This is more of a problem with the relationship between virus and neutralizing antibody concentrations, but practically speaking, inherent variability of the virus input can alter the results up or down depending on how much virus ended up in that assay. That can create more work in trying to put a large dataset together and have the assay data from each run match each other, say by the use of the results of a standard serum/antibody used in each assay.

4) One meta-problem that I've encountered is not the assay's fault, but rather the design of the assay with respect to the target cell. Viruses can enter the cells of various cell types in various tissues by different mechanisms, so the use of a non-physiological cell type for the neutralization assay can give nice, consistent results, but results that aren't useful going forward. For example, the human cytomegalovirus (HCMV) field for years employed primary human foreskin fibroblasts as the cells in the assay for which the neutralization of virus infection by antibodies was measured. This use is/was due to the very limited range of cell types that are available for permissivity of this virus for replication and plaque formation, so the cell type used is one of convenience (which itself is relative since these cells have limited passages). HCMV vaccine development experiments used these cells, and it wasn't for decades was it discovered that viral entry into endothelial cells is a more accurate assessment of neutralizing antibody responses compared to blocking entry/infection in a fibroblast. Thus, the fibroblast data wasn't particularly enlightening in the context of neutralization mediated protection. The cell type in the assay should be scrutinized when embarking on PRNT assays rather than treating them like another reagent in the assay.

I submitted a survey. Best of luck!

Live cell imaging with specific marker proteins

Have a look at this useful link.