Neurobiology 104                           Reading for the next three lectures

October 20, 2003                           G&H Chap. 12 plus pp 231-233

 

 

 

LYMPHOCYTES

 

 

Most kinds of cells belong to discrete "cell types" in which all cells

are equivalent.  In contrast, lymphocytes are extraordinarily

heterogeneous.  Various types of cells that interact with lymphocytes

also are diverse, including reticular cells, dendritic cells, and

endothelial cells.

 

A variety of quite different cell types assume the form of "small

     lymphocytes" when they are inactive.  The small lymphocyte is a

     resting stage for cells circulating in blood or wandering in C.T.

 

4 major classes of lymphocytes are B, T, NK and stem cells.

     Each of these classes is heterogeneous.

 

Classes and subclasses are hard to distinguish morphologically.

 ->  Their most diagnostic features are the proteins on the cells'

     surfaces, called cell differentiation (CD) proteins.

 

Classes are developmentally distinct: e.g. consider B and T cells.

     In birds, pre-T and pre-B cells originate in red bone marrow and

         migrate to separate organs to mature:

     Pre-T cells go to the thymus.  Pre-B cells to the bursa.

         Removing either organ at birth abolishes that class of cells.

     Mammals have a thymus for T cells but no bursa.

         Pre-B cells probably mature in the bone marrow.

 

Lymphocytes do vary in appearance but mainly due to differences in their

states of commitment to divide.

         lymphoblasts --> prolymphocytes --> small lymphocytes

 

Lymphoblasts ("large lymphocytes") have large pale nuclei and

         substantial amounts of faintly basophilic cytoplasm.

         They can actively divide.

Small lymphocytes cannot divide as such.

         They first must dedifferentiate into lymphoblasts.

Prolymphocytes ("medium-sized lymphocytes") are intermediates.

 

WARNING: The "large" lymphocytes of blood are not lymphoblasts.

     Lymphoblasts and prolymphocytes do not circulate in blood.

 

Cells can cycle repeatedly between blast and small lymphocyte forms.


 

 

    A possible killer T cell lineage:

 

                Stage        Morphological form        Location

 

 

                                    Blast          Yolk sac

                                  /

                Stem cell       SL

                                   \               Bone marrow

                                    Blast

                                  /

               Pre T cell       SL                 Blood

                                |

        Thymic lymphocyte       SL = Blast         Thymus

                                |

       Killer type T cell       SL

                                  \

        Stimulated killer           Blast

                                  /

            Memory killer       SL                 Peripheral C.T.

                                  \

      2nd response killer           Blast

 

(SL=small lymphocyte, Blast=lymphoblast)

 

 

                               B cells

 

     B cells can be induced to mature into plasma cells.

         which then secrete massive quantities of antibodies.

     Antibodies are proteins with binding sites for antigen molecules.

 

See G&H Fig. 12-1

 

     A person can make any of 10,000,000 or so types of antibody

         molecules, each with a different amino acid sequence.

 

--> Problem #1: How do genes encode this repertoire of information?

 

    Crucial fact: Each B cell can make only a single type of antibody.

             Antibody diversity comes from diversity among B cells.

 

         A variety of genetic mechanisms generate the enormous

         diversity among antibodies.

              a. Antibody proteins are encoded by multigene families.

              b. The two polypeptide chains of antibodies, L and H,

                 can associate in different combinations.

              c. Antibody genes are split into several segments and

                 each segment is tandemly repeated.  During development

                 one randomly chosen member of each segment is spliced

                 together to form a complete L gene and an H gene.

              d. The exact point of gene splicing is variable.

              e. Extra nucleotides are inserted at junctional

                 sites into the DNA helix in a non-templated manner.


 

 

              f. Point mutations are introduced into the completed

                 L and H genes to produce somatic cell variants.

              g. Other mechanisms:

                    Premessenger RNA's can be differentially spliced.

                    Birds use gene conversion with pseudogenes.

 

B cells differ in their genetic (DNA) sequence from other cell types.

 

--> Problem #2: How do antigens induce only appropriate antibodies?

                (ones which will bind to those particular antigens)

 

        The clonal selection theory proposes a 2-step process.

 

1.  During its formation each B cell generates a unique L and H gene,

        thus irreversibly committing the cell to a particular antibody

        specificity before encountering any antigen.

    It synthesizes a small number of antibody molecules and attaches

        them to its cell surface as a receptor to detect that antigen.

    A trillion lymphocytes wander through a person's blood, lymph and

        connective tissues looking for antigens.  These include cells

        committed to millions of antigens which the person has

        not yet, and in most cases never will, encounter.

 

2. When antigen binds to its receptors, a B cell reverts to a

        blast form and divides into a clone of identical cells.

 

           B cell ----> immunoblast -----> plasma cell

                                           memory B cell

 

      Some progeny cells differentiate into plasma cells.

           (factories for secreting antibody).

      Others differentiate back to small lymphocytes as memory cells

These are easier to reactivate to immunoblasts than before,

and allow a person to respond more rapidly and vigorously

when re-exposed to an antigen (i.e. "booster" response or

"amnestic response").

 

3. A mutation mechanism generates further variation when activated

   lymphocytes are stimulated to divide.

      The mutant cells compete for available antigen.  Those with the

      most avid antibody out reproduce the others.  Thus, the ongoing

      immune response "evolves" towards better and better antibodies.

 

                             T CELLS

 

T cells resemble B cells but are more diverse and complex in function.

 

1. They make a "T cell receptor" protein instead of antibody.

      It: -  is homologous to the antibody molecule.

          -  forms through gene rearrangement.

          -  varies in binding specificity from one T cell to another.

  But it: -  is always bound to the cell membrane.

          -  binds only to antigen which has been digested into small

             peptides that are held on the surface of other cells

  by a "major histocompatibility (MHC) protein.


 

 

2. Subtypes of T cells carry out different functions.

 

 Cytotoxic T cells provide "cellular immunity" distinct from the

         "humoral immunity" from B cells and antibody.

    Rejection of a graft of foreign tissue is an example.

       T cells migrate into the graft, divide, become activated

       as killer T cells and kill the cells of the graft.

         1. by punching holes in the cell membrane.

               They secrete proteins which go into the membrane of

               target cells and assemble into pores.

         2. by inducing the target cell to commit suicide.

               i.e. they induce the pathway for apoptosis.

The main function of cytotoxic T cells is to kill cells that are infected by viruses.

 

 Helper T cells help to activate B cells.

     In general, B cells cannot be activated by antigen unless aided

         by helper T cells which also have been activated.

     T cells also regulate the activation of other T cells.

     Some T cells may regulate formation of other blood cell types:

         eosinophils, mast cells and maybe red blood cells.

 

 Suppresser T cells inhibit the activation of B cells.

     They are important for self tolerance.

         A variety of complex mechanisms prevent a person from

         making antibodies to his own proteins.

     Suppresser T cells can be distinguished from helpers by the

         CD proteins on their cell surfaces (i.e. CD8 vs CD4).

 

3. T cells secrete chemical messengers called interleukins as another

   part of their functions.

 

See G&H Table 12-3

 

4. T cells develop from a complex clonal selection process;

 

   Pre T cells rearrange their T cell receptor gene in the thymus.

        An extra winnowing step in the thymus removes newly formed

        T cells that would react to "self" proteins.

   Cells with acceptable receptors leave the thymus as mature T cells.

 

5. T cells can be activated only by interacting with antigen presenting

   cells (APC's).  These cells:

     - phagocytize extracellular proteins, digest them, place peptide

          fragments on Class II MHC molecules and "present" them on

          their cell surface to T cells.

     - activate T cells whose T cell receptors recognize those

          peptides, using a combination of direct cell-cell contact and

          secretion of chemical messengers.

 

See G&H fig. 12-3


 

 

Antigen presenting cells include dendritic cells, lymphocytes, and macrophages.

 

     T cells also can turn other cells into auxiliary APC's

     by secreting gamma interferon (e.g. endothelial cells).

     Most other cell types of the body also present peptides, but in a

     different way.  They digest samples of their own proteins and

     place fragments on Class I MHC molecules. 

 

Evolutionary considerations:

   1. B cells probably evolved from a subfamily of T cells.

   2. MHC proteins are homologous to antibody molecules.

A large number of other proteins with functions in immunity are homologous in structure to antibodies.

      This immunoglobulin superfamily probably evolved by repeated

        gene duplication and divergence as the immune system became

        more elaborate.

 

 

                         NATURAL KILLER CELLS

 

Natural killer cells (NK) survey the body for tumor cells.  They:

      - correspond to those "large lymphocytes" in blood with

        scattered granules.

      - may actually be relatives of macrophages

      - do not requiring prior induction.

 

There appear to be two or more sorts of NK cells.

 

 

                          STEM CELLS

 

The pluripotential stem cells of bone marrow:

   - can develop into any of the blood cell types.

   - look like lymphocytes when they are inactive.

   - must revert to blast forms to divide.

        The presumed blast form of pluripotential stem cells is

           called a "Hemocytoblast".

        It occurs in bone marrow and in blood.

   - can differentiate into progenitors of individual cell lineages.

   - can circulate in blood as lymphocytes (blast cells do not).

 

A progenitor cell

  -  can give rise to a single cell type.

  -  is usually denoted CFU (colony forming unit), eg CFU-E, CFU-M

  -  can assume the form of lymphocytes at various early stages of

     differentiation into its final cell type.

         eg three early stages in the erythrocyte lineage are

         represented by different lymphocyte forms;

                   CFU-s -> BFU-E -> CFU-E


 

 

Hemopoietic Progenitor and stem cells:

  - are difficult to study histologically because they are rare and

    they all look alike in their blast and resting forms.

  - were identified in cell culture, from the cells they produce when

        activated to divide and differentiate.

  - can be recognized by proteins on their cell surfaces.

  - are activated by secreted proteins called CSF's

    (colony stimulating factors) or poietins (see table 10-6).

 

 

              OTHER POSSIBLE CLASSES OF LYMPHOCYTES

 

Other minor classes of lymphocytes probably await discovery.

    For example, mast cells and dendritic cells originate in bone

    marrow and migrate through the blood to their final location,

    probably in the form of small lymphocytes.

        There are at least two subtypes of mast cells and

        dendritic cells are considerably more heterogeneous.