General Introduction

The Immunology Laboratory course will introduce both classical and contemporary immunological assay procedures within the context of interpreting laboratory science experiments and demonstration fundamental immunological principles.  Your grade will be determined from a variety of evaluation materials including: two formal, research style laboratory reports; two research poster presentations, one submitted as an individual and one as a group project; a laboratory notebook detailing the intent and outcome of the various laboratory procedures; two laboratory exams emphasizing the practical aspects of the procedures; and a subjective assessment of in-class cooperation and performance.  The course is fairly rigorous, requiring consistent effort throughout the semester.  The accompanying laboratory schedule indicates when the individual experiments are conducted and gives the due dates for the reports, presentations and notebook submissions as well as the anticipated times for the exams.  There may be some changes in these dates, but in general this is a reliable compendium of the course chronology.

Each class period identifies a specific experiment to be conducted.  Several of the experiments extend ober multiple laboratory periods.  For purposes of the laboratory notebook, each separate laboratory period should be represented separately.

Three relatively long term experiments will address questions of immune response both in vitro and in vivo.  In groups of four or five, you will conduct these long term experiments following the protocols presented.  You are expected, as individuals, to submit scientific reports on your experimental process and findings.  The first two experiments (Mitogen-stimulated Lymphocyte Transformation and Rabbit Humoral Immune Response) will require submission of a formal laboratory report. The third experiment (Mouse Immune Response) will require submission of "poster presentation" materials as explained below.  Both individual and group presentations will be made; individual poster presentations will be submitted, and following discussion within your group, a final group poster will be prepared for display and discussion.  Experiments conducted throughout the semester, including those for which formal reports and are submitted, will be recorded in a laboratory notebook.  The notebook will be collected twice during the semester.  Two laboratory exams, consisting of both practical and short answer questions about the procedures and their interpretations, will be given during the laboratory period indicated.  Unannounced quizes on laboratory objectives will occur occasionally throughout the semester.  (Exams and quizes will total 170 pts.)  An additional 60 points for laboratory performance (measured largely by participation and cooperativity) completes the 500 total points for the laboratory grade.

You must get 55% of the total points (275 our of 500 offered) in order to pass the class; turning in all the required work will generally insure that goal.  Written work which is turned in late will be docked 10% of each school day it is late, that is, work that is two weeks overdue has 0% value.  The final grade distribution is curved based on clearly separated groupings of students, that is students separated by a few percentage points overall will generally receive the same grade.  In general, students who accumulate 85% of the points or better receive A's, 75%-84% receive B's, 64%-74% receive C's, 55%-64% receive D's, and less that 55% receive F's.  Incomplete grades are issued for chronic medical reasons.

LABORATORY NOTEBOOK (100 points):

An organized arrangement of the laboratory experiments performed throughout the semester.  It should include a Table of Contents to aid in the organization.  Other inclusions are : the laboratory instructional handout; notes on specific procedures and any modifications of the handout instructions, intended or not; observations and suggestions to help you if you have to do this again; experimental results in numerical counts, observations or drawings (appropriately labeled), any or all that are appropriate; comments on the experience.

LABORATORY ATTENDANCE/PARTICIPATION (60 points):

Grading in the category will represent an assessment of preparation and participation in experimental procedures throughout the semester.  Laboratory attendance is required.  Unexcused absences cost 25 points each.  There are a number of group experiments in the laboratory; in some experiments, students will be required to perform out-or-class procedures for their particular group; within scheduling limitations, shared participation is expected.

FORMAL LAB REPORTS (120 pts):

Style is to be standard research paper format. It includes: (1) an Abstract or Summary of 250 words or less which briefly explains or outlines the purpose of the experiment, the general procedure, the results obtained, and conclusions to be drawn from the experiment; (2) an Introduction which describes the purpose of the experiment with background information about the procedures; (3) a Materials and Methods section which should briefly describe what you did, not by simply copying the laboratory notes, but summarizing the types of procedures e.g. if the laboratory notes say, "Centrifuge the blood at top speed on the clinical centrifuge and remove the upper layer (serum) component to a clean tube", you say something  like "Serum was isolated from the blood by centrifugation"; (4) the Results section should detail what you observed and/or include any graphs you made, and it should be presented in a fashion that facilitates the reader's understanding of what experiments were undertaken and what the outcome was of those experiments; (5) the Discussion and Conclusions section should attempt to explain the results in light of current knowledge, i.e. were the results obtained those that were expected and how can you explain any discrepancies? (6) Bibliography of cited material, at least one outside source is required.  An article on science report writing is available, on reserve, in the library.

POSTER PRESENTATION: (50 pts-25 individual; 25 group)

For purposes of this course, a poster consists of materials appropriate for a formal presentation of work completed.  Rather than writing a laboratory report for the mouse immune response experiments, you will be required to submit a mock poster.  You will have to analyze the data obtained and organize it for presentation into figures, graphs or tables with appropriate legends.  A student of your educational level who has not conducted this experiment should be able to interpret theses presentations; visual clarity of what you are trying to demonstrate should be evident and will represent a major component of the grading.  Accompanying this collection of figures, etc. should be a complete ABSTRACT, analogous to those prepared for formal reports, summarizing the information necessary for interpretation of all aspects of the experimental process, i.e., introduction, methodology, results, discussion and conclusion.  Concise statements for an Introduction, Methods and Discussion/Conclusion, no more than one page for each section, will serve as the rest of the presentation.

LONG-TERM EXPERIMENTS:

Lymphocyte Transformation

Antigenic stimulation of select lymphocytes is the key event in specific immune responses.  Stimulated lymphocytes undergo morphological and physiological changes, termed blast transformation, which can be monitored both microscopically and metabolically.   We will monitor lymphocyte transformation in vitro, i.e. in cell cultures, using compounds which stimulate large populations of lymphocytes to undergo "clonal expansion".  The response to these compounds will be assessed morphologically by phase contrast microscopy and metabolically both by observing carbon dioxide Influences on the culture media and by incorporation of radioactive thymidine into the DNA of mitotically active cells.  The experiment will take two weeks to complete and will require a laboratory report worth 50 points.

Rabbit Humoral Immune Response

A long term experiment will be conducted during the semester on the immune response of laboratory rabbits to injection with an immunogen, chicken ovalbumin.  The response will be monitored following two inoculation procedures: one with the ovalbumin alone, and another with ovalbumin accompanied by adjuvant, an immune response enhancing material.  These immune responses will be studied as they develop over nearly half of the semester.  Thus the laboratory portion of the course will start with this experiment in order to provide adequate time for the response to develop, and will include periodic bleedings from the rabbits in order to obtain blood samples containing the serum antibodies for assessment.  Assessment of the levels of antibody production will be determined using two assay procedures: ELISA and Dot Blot.  The entire experiment will take approximately seven weeks and will require a formal lab report worth 70 points.

Mouse Cellular Immune Response

White Blood Cell activities are fundamental to immune responsiveness and this lab report will focus on these cellular activities.  This experiment involves a study of the in vivo immune response Of laboratory mice to the injection of an immunogen, sheep red blood cells (SRBC).  The mice will be inoculated with SRBC through one of two routes:  intraperitoneally or intravenously.  White blood cells will be isolated from the spleens of the inoculated mice in order to determine whether specific cellular effects directed against sheep red blood cells have developed.  One assay procedure, Rosette Formation, will be used to assess the immune response for development of cells which directly interact with the SRBC.  The entire experiment will take approximately one week to complete and will require submission of a poster presentation worth 50 points due in two parts, once as an individual and once as a group.