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46 Cards in this Set
- Front
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3 important immunoglobulin protein types |
Monoclonal Polyclonal Oligoclonal |
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autonomous, ag-independent transformation of a homogenous single clone of B cells. Paraprotein or M-component restricted band of mobility on electrophoresis Ig restricted to one heavy chain and/or one light chain secreted in large quantities Found in malignant, benign, or transient disorders |
Monoclonal Immunoglobulin |
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the increased production of different classes of Igs due to the transofrmation of many clones of B cells Can be observed in: ~Acute and chronic infections ~AIDS ~Sacrocoidosis ~Connective Tissue and liver diseases its what we expect to see |
Polyclonal Immunoglobulin |
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What is the darkest bank on a SPE |
Albumin |
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restricted electrophoretic mobility in agarose gel >4 bands in CSF ONLY, positive Detected in the gamma region, produced in CNS Disease States: ~viral infecttions ~meningitis ~neurosyphillis ~lupus ~D. melitus ~post bone marrow transplant |
oligoclonal immunoglobulins |
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what kind of samples do you need for the oligoclonal |
serum and CSF |
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the pH at which a particular molecule carries no net electrical charge |
Isoelectric point |
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isi the method used to detect oligoclonal bands in patients with Multiple Sclerosis |
Isoelectric focusing |
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Proteins are + at pH values ____ their pI |
below |
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Proteins are - at pH values ____ their pI |
above |
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the gel is a pH gradient pH gradient+influence of an electric field= protein move to where the gradient is net zero If a protein diffuses away from its pI, it gains charge and migrates back This is the focusing effect of ____ which concentrates proteins at their pIs only oligoclonal |
IEF
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what allows proteins to be separated on the basis of very small charge differences |
IEF |
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Demyelinating, inflammatory disease CNS Ab directed against a myelin basic protein peptide Ig increased in spinal fluid (~90 pts) multiple plaques on brain |
Multiple Sclerosis |
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compares amount of IgG in spnal fluid to that of serum quantitative measurement |
IgG index |
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testing aids in diagnosing MS |
IgG and Oligo |
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an abnormal clonal proliferation of transformed plasma cells that leads to overproduction of a single monotypic Ig 3 pathogenic signs: 1. B cell clones with no apparent Ag stimulus 2. Monoclonal bands 3. Abnormal levels of protein in the blood |
Monoclonal Gammopathy |
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Disease States of Monoclonal Gammopathy |
Multiple Myeloma Waldenstrom's Macroglobulinemia Amyloidosis |
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the uncontrolled proliferation of a single clone of plasma cells typically present in the bone marrow Clinical Findings: ~weakness, anorexia, and weight loss ~bone involvement with skeletal destruction ~Pain, anemia, and renal insufficiency ~various neurologic deficits ~recurrent bacterial infections |
Multiple Myeloma |
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Produces excess homogeneous immunoglobulin or immunoglobulin fragment immunoglobulin produced is usually IgG sometimes IgA, IgM, and kappa or lambda only |
Multiple Myeloma |
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The unbalanced synthesis of light chains not incorporated into intact Ig molecules SPE usually shows a markedly reduced gamma region w/o a spike or band The free light chain is rapidly cleared from the blood by glomerular filtration into urine cant be detected by serum IFE, because is more sensitive that SPE |
Light-chain Myeloma Bence Jones Proteins |
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polyclonal (normal) free light chinas in urine that migrate as 3-7 equally spaced small bands with equal intensity |
Ladder Effect |
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is a monoclonal light chain, due to a plasma cell dysrasia (unspecified blood disorder) |
Bence Jones Proteins not Ladder |
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a malignant proliferation of IgM-producing plasma cells Plasmacytoid lymphocytes infiltraate the bone marrow, spleen, and lymph nodes known for its "sheets of plasma cells" on bone marrow slides |
Waldenstrom's Macroglobulinemia |
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weakness, ease of fatigue, headache, and weight loss as order progresses patients present with hyperviscosity syndrome (fluids resist to flow) |
Waldrenstrom's Macroglobulinemia |
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a spike in the beta or gamma region on SPE IgM is increased, IgG and IgA decreased Serum IFE= monoclonal IgM kappa or IgM landa protein relative serum viscosity is elevated |
Waldrenstrom's Macroglobulinemia |
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rare & potentially fatal disease characterized by deposition of a protein in tissues amyloid protein-produced by cells in bone marrow and are deposited inany tissue or organ found association with Multiple Myeloma Lamda is more common than kapp |
Amyloidosis |
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3 stages of amyloidosis |
Primary Secondary Familial |
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this amyloidosis stage may occur in association with multiple myeloma |
primary |
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this amyloidosis stage is a result of chronic infection or inflammation (TB or arthritis) |
secondary |
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this amyloidosis stage is produced from an inherited varient protein |
familial |
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Symptoms: swelling of ankles and legs weakness significant weight loss shorntness of breath numbness or tingling of hands and feet severe fatigue skin changes such as thickening or easy bruising purplish patches around the eyes enlarged tongue |
Amyloidosis |
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presence of monoclonal immunoglobulin in serum and/or urine, but no evidence of malignancy |
Benign monolconal Gammopathy |
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2 distinct monoclonal immunoglobulins (IgG Kappa and IgM kappa) but is RARE |
Biclonal Gammopathy |
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Serum/urine transferred to support mediium, an electrical charge is applied protein is separated into 5 zones (charge/migration) proteins have a neg charge and migrrate toward the annode Gel stained, scanned, and evaluated |
SPEP |
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What are the 5 zones |
albumin alpha-1 alpha-2 beta gamma |
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sensitive method that achieves better resolution of the proteins beyond the classical five band patterns is accomplished by : ~agarose gel ~high voltage coupled with a cooling system ~electrophoretic buffer |
High Resolution Electrophoresis (HRE) |
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which HRE protein fractions go with which zone 1. pre albumin, albumin 2. alpha-lipoprotein, alpha-1 antitrypsin 3. alpha-2 macroglobulin, haptoglobin 4. Transferrin, beta-lipoprotein 5. Fibrinogen, Igs, CRP |
1. albumin 2. alpha-1 3. alpha-2 4. beta 5. gamma |
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useful inn identification of IgG-specific oligoclonal banding in paired serum and CSF samples the most sensitive method and 2X as sensitive as IgG index is used as an aid in the dx of inflammatory disease of the CNS, such as MS |
Isoelectric Focusing (IEF) |
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serum and CSF proteins are separated according to their isoelectric points (pI) in an agarose gel when a direct current is applied, a pH gradient is established in the support medium Proteins are transferred to nitrocellulose, which is fixed to visualize IgG specific bands the patter are interpreted qualitatively by comparing the presence or absence of OLG bands in serum and CSF |
IEF Principle |
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The OLG bands resolved are what type of immunoglobulins |
IgG |
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Oliogclonal banding in CSF in IEF but not in what |
serum |
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combines SPEP, Immune precipitation, double diffusion aids in the dx and evalutation of the therapeutic responce in many disease states affecting the immune system ususally indicated when SPEP has indicated a rise at the Ig level |
IEP |
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2 stage procedure using agarose gel electrophoresis in the first stage and immunoprecipitation in the second |
IFX |
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specific antisera is added to a template that is placed over the electrophoresed areas whenever the pts immunoglobulin reacts with antisera, a precipitate will form in the gel the gel is then stained with acid violet observed for bands of restricted mobility |
IFX |
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Advantages of IFX over IEP |
20x more sensitive much easier to interpret |
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Factors that infuence Rx in Gels |
+/- charge of protein (main) pH (8.6 is best) Temperature (incr= faster diffusion) Ionic strength (higher ionic s.=less diffusion) Support/medium |
