MBI - 414 Immunology Principles
MBI - 415 Immunology Principles and Practice

Immuno FAQs


Dropping or Withdrawing from Courses - Complete Version

The full text of the new withdrawal policy that goes into effect this term, excerpted from Registration Proceedure: Sections 01.203.E and 01.203.F in the Student Handbook, is reproduced here:


Interesting Q&A from Previous Classes ... and others

Q: How important is it to know the names of the various people presented in your outlines. There are an awful lot of people, and I am struggling trying to remember specific people and the specific activity that they did.
A: There will be at least one question that relates to major advances in the field of immunology and the names of the people involved will, at the very least, be important clues to which advance you need to discuss. My aim in presenting the experiments and telling you who did them is to help you see how the field developed.

Q: I am a PhD student at Iowa State. I was just browsing and chanced on your immunology class notes. I have a question on antigen-antibody interactions. In specific antigen-antibody interaction for the formation of the various noncovalent bonds how much time is needed? Or in other words what is the time scale of these interactions?
A: Formation of the initial interactions between antibody binding sites and the antigenic determinants for which they are specific is diffusion rate controlled (sensitive to variations in temperature, viscosity and, molecular weight, especially in gels). Therefore, they occur on the time-scale of seconds, perhaps faster (since the experiments to determine this were done a long time ago (Heidelberger and Kendall. 1935. J. Exp. Med. 61:563-591) and the sophisticated equipment we have now was not available to do the reactions more quickly). Formation of stable precipitable antigen-antibody complexes for soluble antigens is, however, a two-step process, and the second step takes hours (we usually incubate 1-2 hr at 37C, then overnight at 4C to allow for this). December 9 Q: I was wondering if B7.1 and B7.2 were just parts of the whole B7 molecule that interacts with CD28, or if only one portion of the B7 (like just B7.1) interacted with CD28?
A: If you check out page 39 of your textbook, you will see that B7.1 and B7.2 are alternative forms of B7

Q: When we discussed the spleen, I think that I may have interpreted your drawing wrong. The notes I took indicate that antigen travels through the splenic artery and is dumped into the red pulp where it encounters a dendritic cell and is presented to a T-cell which releases cytokines and triggers a b cell to divide and differentiate into ab producing plasma cells and memory cells. My confusion is that figure 2-19 in the book shows the red pulp surrounding the B-cell rich marginal zone, and the T-cell rich PALS at the center of the spleen. So does antigen get dumped into the red pulp, taken up by a dendritic cell, then the dendritic cell travels through the marginal zone to the PALS(in the center) and presents it to a T cell and the T cell travels back out to the marginal zone to release cytokines to the B cells? I am confused about the path of this process.
A: The antigen diffuses from the red pulp into the white pulp, where it:
1) gets phagocytized, processed and presented to T cells
or
2) diffuses into the B cell rich area of the PALS (follicles) and is bound to BCRs with appropriate specificity.

Q: Okay, the first question is based on antibody/antigenic binding. For some reason I had thought that the binding of these two was a collection of noncovalent interactions, but in the book it describes the affinity labeling experiment as AA covalently bonded to the residues on the antibody binding site. If they are covalent, does that make this question false: Using affinity labeling techniques, Kabat (and others) showed that the hypervariable regions of antibody molecules contain contact residues (amino acids that actually form NONCOVALENT bonds with the antigenic determinants) and are therefore considered to be the CDRs.
A: The way all antibodies interact with determinants is via noncovalent interactions. Kabat and others capitalized on this to localize their ligands in antibody combining sites. Once the ligand was bound to the combining site, they photoactivated a reactive center in the ligand so it would also form a covalent bond with one of the amino acids in its vicinity. The assumption that an amino acid that was located close to (within?) the three-dimensional location of the bound ligand was one that was involved in binding the ligand was the basis for the idea that this experiment would determine the location of the CDRs.

Q: Okay, the next question I have is just nomenclature type... The effector cell of cytotoxic cells is CTLs right? then what do just plain old Tc cells do? are they merely a step in the maturation of CTLs?
A: Tc are the precursors of CTLs ... Tc cells will become CTLs when they are activated.

Q: In the notes I have Neutrophils, Basophils and Eosinophils as being in the bone marrow and only mast cells in the tissues, yet in the book they often talk about them migrating from the blood to the tissue?? do these become mast cells when they reach the tissue?
A: The granulocytes (Neutrophils, Basophils and Eosinophils) are all cells found primarily in the blood. They do, however, migrate into the tissues during inflammatory episodes ... where they die within a couple of days. In contrast, mast cells do not mature until their precursor cells migrate into the tissues ... and they live in the perivascular tissues for years (not just hours-to-days).

Q: B cells bind to soluble antigen right? does this mean that if an antigen is insoluble, that B cells will not recogonize it at all? and thus they cannot become APCs?
A: B cells can bind to either soluble or insoluble antigen. They don't "care" about it's form, just as long as there are determinants available for their BCRs to bind in a specific manner.

Q: i just had a quick question about T cell response, i understand why T cells would respond to denatured antigen because they recognize antigen that is processed into antigenic peptides, but don't B cells typically present the antigen.. so if the antigen was denatured, how does the MHC get on the antigen for the T cells to respond.
A: You're right when you say that B cells can present antigen to T cells. However, they only do that after they have specifically recognized the native form of the antigen via their BCR (the antibody molecules on their surface). BCR recognition is what we refer to when we say that B cells only respond to native antigen ... since native antigen (whatever is present in the extracellular fluids) is what they bind with their BCR and that is what stimulates them to make and secrete antibody molecules. Binding of processed antigen to MHC-2 for presentation is only done during antigen processing ... when partially degraded antigen is being generated for that purpose. That's not a response to the antigen per se.

Q: The figure with the apple and the 6 fingers around it that you showed in class I had some questions to clear it up. Is the apple represent the antigentic determinats?
A: The apple represents a single antigenic determinant.

Q: ... and 3 of the 6 fingers represent the light chain complementarity determing regions, and the other 3 represent the heavy chain complementarity determing regions?
A: Yes, that's exactly right!

Hello. I was reading about the different classes of antibodies, and I have a few questions about IgE.
Q: In humans, does IgE play a role in any immune response besides hypersensitivity associated with allergic reactions?
A: Yes ... it is involved in mediating immunity against anti-parasites (intestinal protozoa, worms, etc.). It binds to surface receptors on eosinophils and "arms" them to deal with these parasites.

Q: Is IgE a more prominent antibody type in other species of animals?
A: Other animals have immunoglobulins that serve the same purposes, but most do not have Ig molecules with similar enough structure to be called IgE.

Q: What advantages (if any) does IgE bring to an organism?
A: See the answer about arming eosinophils (above).

Q: Since it is significantly less abundant than the other classes and it is responsible for harmful allergic reactions, would an organism potentially be better off without it?
A: It is actually not less abundant than the other Ig classes ... unless one only looks in the fluid phase of the blood. Remember, it is mostly found attached to surfaces of basophils and mast cells throughout the body. People who have few or no intestinal parasitic infections might actually be "better off" without IgE ... as long as they could guarantee that they would never get these infections. It appears that IgE-mediated allergies in developed countries are such a problem precisely because we don't have the parasites to contend with. It's a long story (and it's mostly conjecture) but the "gist" of it is that people who have few or no intestinal parasitic infections are destined to have more type-I hypersensitivities (than those who have the infections) because most of their IgE antibody molecules are specific for allergens, rather than parasites. To trigger histamine release from basophils or mast cells (via IgE mediation), you need two adjacent surface IgE molecules with ability to bind to determinants on the same allergen molecule. If you think about the probability of this happening, you can immediately see that "diluting out" the anti-allergen IgE molecules with anti-parasite IgE molecules (i.e., increasing the specificity heterogeneity of the IgE population) would decrease the tendency to get allergic reactions when exposed to the allergen after sensitization. How about that?

Q: I'm still having trouble with the idea that MHC class 1 molecules present our own proteins to other cells and can't seem to understand how this brings on an immune response. Are these not host proteins no matter how you look at them. It shouldn't matter if a virus is "telling" us to make it, it is still a host protein. Maybe it's that we would not normally present these proteins on our cells?
A: Presentation of peptides derived from our own proteins on our MHC class 1 molecules does *not* bring on an immune response. This is because our own proteins are not recognized - i.e., they are not foreign. The key to foreignness is *not* being present in the thymus during development of T cells. When T cells are formed, we first develop a large population of T cells representing a very broad spectrum of TCRs, then we eliminate all those cells whose TCRs recognize processed antigens being presented to them by APCs in the thymus. The antigens that are presented during this time are self antigens (provided the host is not infected at the time). So . . . the only T cells able to respond are those that recognize things that were *not* present while the T cells were being formed, and these are non-self proteins. Peptides from a viral protein (one that was not being synthesized during development of T cells) would, then, be recognized and an immune response would be induced.


Q: How exactly does photo affinity labeling work and what does it do?
A: Photo affinity labeling is a technique used to determine which amino acids in an antibody molecule are contact residues in antigen-binding sites of antibodies.
It is done via these steps:

Overall, this approach was used to show that (a) the variability in antibody molecules (as one looks at molecules with differing specificity) occurs in three restricted areas of the N-terminal ~110 amino acids of both light and heavy chains, and that (b) this variability is in the same areas (amino acid residues) that bind antigenic determinants (i.e., are contact residues).


Q: While preparing for this upcoming exam, I am primarily using the web notes and in class notes to study from and using the text as a secondary resource for any information I don't understand. concepts that you breifly cover in lecture and that the book goes in depth about, are we required to know the depth of the book or your lecture?
A: I expect you to know the information I present in class at the depth I present it, not more. However, the notes on the web are not as detailed as the lectures themselves; they are intended to be study guides, not the complete story.

Q: When antibodies assemble, how do heavy chains know to bind another heavy chain with the same light chain attached?
A: When antibodies assemble, heavy chains bind another (identical) heavy chain with the same light chain attached because:

Q: If the ITAMS in BCRs and TCRs are not attached to the BCR (TCR), how does it know that the ITAM was phosphorylated and to start producing antibodies?
A: Phosphorylation of the ITAMs is the first step in a rather complicated sequence of events that ultimately results in a signal to the promoter region of the genes for antibodies (in B cells) or for cytokines, etc. (in T cells). The BCR or TCR never gets the signal . . . it is only instrumental in initiating it (by helping the ITAMs get phosphorylated in the first place).

Q:
What is the best way to study for this course?
A: Check out the Study Tips and do what they tell you. The tip about using writing to integrate diverse ideas and synthesize them into your own working model is my favorite. Another very helpful approach is to thoroughly familiarize yourself with the material, then discuss it with other students to help you look at it from different viewpoints to make sure you understand it and that you have not just memorized the information.

Q: Exactly how useful is the textbook to us in studying for this class?
A: The textbook is a very important reference, because it will allow you to go beyond the lectures and the study guide on many topics, thus allowing you to better develop your knowledge base and your understanding of microbiology.
As I alluded to in the Study Tips page, it would be a mistake to think that memorizing your lecture notes is sufficient for success in this course. After you have memorized enough material to have a significant knowledge base, you need to use it as a "jumping-off" point for developing a deeper understanding of immunology, especially from a conceptual and process-oriented viewpoint. Only when you have mastered this process will you be able to truly understand immunology to the degree that you can use it for the rest of your life. That, of course, is what a real education is all about.
Interestingly enough, as you progress through the semester, one way to gauge your progress is to note how much easier it becomes to read both the textbook and, especially, journal articles.
Happy synthesis and integration!

Q: What will the questions be like on the exams?
A: Check out the Sample Questions for some tips and mind tweaks.

Syllabus

Laboratory

Instructor

Study Guides

Sample Questions

Cool Immuno Stuff

Study Tips

Immuno FAQs

Bugs'n'Drugs


© 1996-2011 John R. Stevenson. All Rights Reserved

Please
email questions and comments to:
John R. Stevenson, Ph.D.
Associate Professor
Department of Microbiology
Miami University
Oxford, Ohio 45056
USA
This document was last modified on Saturday, 20-Aug-2011 17:47:31 EDT