Lecture 10 (labs) -Exam 5 Flashcards

Question

RNP * What is this? * When does antibodies to RNP occur?

Answer 1

* A small nuclear protein * Antibodies to RNP occur in approximately 50% of patients with Lupus and in patients with connective tissues diseases- specifically mixed connective tissue disease

Answer 2

Mixed connective tissue disease is characterized by high levels of RNP antibodies without detectable SM or ds DNA antibodies What is a mixed connective tissue disease? * Diagnosed with positive RNP and there are features of at least two connective tissue disease, including systemic lupus erythematosus, systemic sclerosis, polymyositis, dermatomyositis, and RA

Answer 3

* Use to identify patient with multiple myeloma and other serum protein disorders * Electrophoresis separates proteins based on their physical properties and the subsets of these proteins are used in interpreting the results * The serum is placed on a specific medium and a charge is applied * The pattern of serum protein electrophoresis results depends on the fraction of two major types of protein- albumin and globulins

Answer 4

* Laboratory tests that measure different parts of the immune system are important for diagnosis someone with primary immunodeficiency and determining which of the over 450 different conditions they may have * **Basic panel of immune function tests: CRP, IgM/IgG, IgA, ANA, total protein**

Answer 5

* Antibody deficiencies * Cellular (T cell) defects * Neutrophil disorders * Complement deficiencies

Answer 6

* To test specific antibody production, the person is immunized with common vaccines, and blood samples are obtained immediately prior to and approximately four weeks after the immunization to evaluate how well the individual forms specific antibodies. * It is worth noting that during the maturation of the immune system, the response to carbohydrate antigen vaccines lags behind the response to protein antigen vaccines. This is the reason for having a childhood version of the pneumococcal vaccine, the PCV13, which makes it easier for infants to respond to Streptococcus pneumoniae.

Answer 7

* Vaccines provide protection by inducing humoral and/or cellular immunity to the pathogens causing disease. With respect to humoral immunity, the dense surface distribution of often unique glycan structures on diverse pathogens and on malignant cells makes carbohydrates attractive vaccine targets * **It is now well known that immunization with neoglycoconjugates composed of capsular polysaccharide-derived glycans covalently coupled to an immunogenic protein carrier (conjugate vaccines) induces long lasting protection against encapsulated bacteria even amongst persons in high risk groups**

Answer 8

* To find out if you have had a recent or past infection * Check out vaccination status – If your medical records are incomplete and you don’t know your vaccination status * To find out if a vaccine is effective - the test shows if your previous vaccine is providing enough protection * Requirements for many jobs and or school admission – to show proof of vaccination

Answer 9

* For the test, **a patient’s blood sample is diluted serially and incubated with known antigens to determine the presence of antibodies against these antigens**. The number of dilutions to be prepared depends on the concentration of antibodies in the blood. * A higher number of dilutions are needed for a sample wherein the antibody concentration is very high. **The test measures the highest dilution of the blood at which antibodies cannot be detected anymore** * The result of an antibody titer is generally presented as a ratio. **For example, a ratio of 1:200 indicates that diluting one part of the blood sample to 200 parts of the diluent solution (saline) finally leads to an undetectable antibody level in the blood sample**

Answer 10

A successful antibody titer result depends on the type of antibody being detected. * immunoglobulin M (IgM) appears in the blood between 2 – 4 weeks post-infection, * whereas immunoglobulin G (IgG) takes around 4 – 6 weeks

Answer 11

IgM antibodies are the first immunoglobulins your body makes after you're exposed to germs. They provide short-term protection while your body makes other antibodies. IgM antibodies are in your blood and lymph fluid

Answer 12

IgG antibodies are very important for fighting infections from bacteria and viruses. Most of the immunoglobulins in your blood are IgG. You also have some IgG antibodies in all your body fluids. Your body keeps a "blueprint" of all the IgG antibodies you have made. That way, if you're exposed to the same germs again, your immune system can quickly make more antibodies.

Answer 13

IgA antibodies protect your respiratory tract and your digestive system from infections. You have IgA antibodies in your blood, saliva, and gastric juices.

Answer 14

An immunoglobulins blood test

Answer 15

* The **standard screening test for humoral immune** function starts with measuring immunoglobulin levels in the blood. These consist of IgG, IgA, and IgM….sometimes IgE * There are also tests to measure the different types of lymphocytes in the blood, specifically B cells which produce antibodies * **B cells may be absent in certain antibody deficiencies such as X-linked agammaglobulinemia (XLA)**

Answer 16

Cellular T cell immunity- determines the numbers and different types of T cells * **First time this is tested is at birth as part of the newborn screening tests** * Can be checked with flow cytometry * Can also be looked at within the CBC with diff as the absolute lymphocyte count (75% of circulating lymphocytes are T cells)

Answer 17

Most common primary immunodeficiency characterized by undetectable serum IgA * Have no common mendelian pattern, can also be caused by medications, certain viruses

Answer 18

This class of antibodies is composed of four different subclasses: IgG1, IgG2, IgG3, and IgG4. * Each subclass is present in different concentrations in the blood and those concentrations vary with age. * The IgG in the bloodstream is 60-70% IgG1, 20-30% IgG2, 5-8% IgG3, and 1-3% IgG4. IgG1 and IgG3 reach normal adult levels by 5-7 years of age, while IgG2 and IgG4 levels rise more slowly, reaching adult levels at about ten years of age.

Answer 19

* Each subclass serves predominantly (but not exclusively) a slightly different function in protecting the body against infection. For example, IgG1 and IgG3 subclasses protect against toxins from bacteria such as diphtheria and tetanus, and against viral infections. * In contrast, IgG2 predominantly protects against the polysaccharide (complex sugar) capsule of certain disease-causing bacteria such as Streptococcus pneumoniae and Haemophilus influenzae.

Answer 20

Individuals with IgG subclass deficiency are defined as having recurrent infections, persistently low levels of one or more IgG subclasses, and normal concentrations of total IgG, IgM, and IgA. * IgG2 or IgG3 deficiencies are the most common IgG subclass deficiencies. Since IgG1 comprises 60% of the total IgG level, having a deficiency of IgG1 usually drops the total IgG level below the normal range, resulting in hypogammaglobulinemia. * IgG2 or IgG3 deficiencies would show a normal total IgG since IgG1 is the most common in blood

Answer 21

* **IgM is the largest in size of all immunoglobulins** - it contains five antibody molecules held together by the J chain. IgM antibodies serve as a first line of defense against bacteria, viruses, and fungi and in protection against autoimmune diseases and inflammation. * It is predominantly present in the circulating blood versus IgG, which is present both in the circulation and in the tissues, and IgA, which is predominantly in secretions. * IgM is the first immunoglobulin secreted during an initial exposure to infectious organisms or a vaccine. IgM is also different from other immunoglobulins in that

Answer 22

* **Although described more than 50 years ago, it is only recently that selective IgM deficiency has been included in the list of forms of PI.** * It is characterized by low (partial) to absent IgM (complete) in the blood but normal IgG and IgA levels. It occurs equally in children and adults and in both men and women. * **Partial selective IgM deficiency is more common** than previously realized, whereas complete selective IgM deficiency is rare.

Answer 23

Individuals with selective IgM deficiency may be asymptomatic or symptomatic. Of those who are symptomatic, approximately 80% present with predominant bacterial infections. * Among infections, the most common are chronic sinusitis, upper respiratory tract infections, bronchitis, and pneumonia. **Almost 40% of individuals with selective IgM deficiency have allergic diseases, including hay fever and asthma.**

Answer 24

Almost 40% of individuals with selective IgM deficiency have allergic diseases, including hay fever and asthma. Autoimmune diseases are observed in about one-third of all individuals with selective IgM deficiency. Autoimmune diseases are more common in adults than children with selective IgM deficiency. * Example: SLE, RA, and autoimmune thrombocytopenia, Hashimoto’s thyroiditis and Addison’s disease

Answer 25

* if the serum IgM level is below two standard deviations of the mean for age-matched controls. * in children, should only be established after months of follow-up. * To confirm a diagnosis of primary selective IgM deficiency, other possible causes of IgM deficiency must be excluded, such as leukemia, lymphoma, and immune suppressive medications, such as Clozapine.

Answer 26

Neutrophils play an essential role in immune defenses because they ingest, kill, and digest invading microorganisms – first line of cells recruited at the site of infection * Normal WBC count 4000-11,000 cells/microL, out of which 60-70% are mature neutrophils circulating in the blood * Failure to carry out that role leads to immunodeficiency characterized by the presence of recurrent infections

Answer 27

* Defects can be **quantitative- neutropenia or qualitative- neutrophil dysfunction** * Can obtain a CBC with differential that shows the absolute neutrophil count * Must look at a blood smear to rule out certain diseases that are associated with abnormalities in the structure of neutrophils * Sometimes requires a bone marrow biopsy to see if neutrophils are being made properly * Assess in patients with lymphoma undergoing chemotherapy

Answer 28

with high morbidity and mortality

Answer 29

Just know that we can calcuate it

Answer 30

**Neutrophilia is defined as a higher neutrophil** count in the blood than the normal reference range of absolute neutrophil count * Absolute neutrophil count (ANC) typically ranges between 2500 to 7000 neutrophils/microL * **Neutrophilia is defined as an absolute neutrophil count greater than 7000 neutrophils/microL**

Answer 31

**infections**, inflammation, and/or neoplastic processe

Answer 32

* Normally, there is a balance between different types of white blood cells, including neutrophils, lymphocytes, monocytes, eosinophils, and basophils. Neutrophils are the most abundant type of white blood cell and play a crucial role in the body's defense against infections. * **In a left shift, there is an increase in the number of immature or "band" neutrophils circulating in the bloodstream. These immature neutrophils are released prematurely from the bone marrow in response to an increased demand for white blood cells, typically due to an acute infection or inflammation.** ## Footnote The term "left shift" originates from the graphical representation of the CBC results, where the different types of white blood cells are arranged from left to right based on their maturity.

Answer 33

Acute and serious infections * start antibiotics

Answer 34

* The standard screening test for deficiencies in the classical complement pathway is the total hemolytic complement assay or CH50 * Specialized complement laboratories can provide additional testing that will identify the specific complement component that is defective

Answer 35

* **Decreased levels of complement arise from immune-complex disorders** such as SLE and other selected forms of vasculitis * NOT useful for screening of SLE, but it often used to monitor disease activity in patients with SLE

Answer 36

the bone marrow of the skull, ribs, sternum, vertebral column, pelvis, and femurs

Answer 37

* Lymphocytes represent around 20-40% of WBC’s * Relative lymphocytosis is an increase in WBC’s of more than 40% in the presence of a normal absolute white cell count

Answer 38

Low white blood cell count * Primary Lymphopenia: is rare and often results from genetic or developmental abnormalities that affect the production, function, or survival of lymphocytes. Examples include severe combined immunodeficiency (SCID), DiGeorge syndrome, and other primary immunodeficiency disorders. * Secondary Lymphopenia: Secondary lymphopenia is more common and can be caused by a variety of factors

Answer 39

* **Infections**: Viral infections (e.g., HIV, Epstein-Barr virus, cytomegalovirus) can directly affect lymphocyte production and function. * **Medications**: Certain medications, such as corticosteroids, chemotherapy drugs, immunosuppressants, and antiretroviral therapy, can suppress lymphocyte production or induce cell death. * **Autoimmune Disorders**: Autoimmune diseases, such as systemic lupus erythematosus (SLE) and rheumatoid arthritis, can lead to lymphocyte destruction or dysfunction. * **Hematologic Disorder**s: Conditions affecting the bone marrow, such as aplastic anemia, myelodysplastic syndromes (MDS), and leukemia, can impair lymphocyte production. * **Radiation Therap**y: Exposure to ionizing radiation, either as medical treatment or environmental exposure, can damage lymphocytes and suppress immune function.

Answer 40

* High white count * **Need to distinguish between reactive and malignant** * Is defined as an absolute lymphocyte count over 4000 cells/microL * Lymphocytosis is detected incidentally in the majority of the patients. * The evaluation starts with an extensive history and physical and review of peripheral blood smear review, which may direct for the need for further testing (flow cytometry, karyotype, and FISH).

Answer 41

Clues to the presence of clonal lymphoproliferative disorders include a highly elevated ALC, presence of abnormal lymphocyte forms on PBS, and the presence of other cytopenias.

Answer 42

EBV, CMV, Acute HIV infection (chronic HIV causes lymphopenia), influenza, measles and mumps, adenovirus

Answer 43

* Bartonella henselae – leads to cat scratch disease – lymphocytes are large and atypical * Bordetella Pertussis – increased lymphocytes that are small with a deeply cleaved nucleus * Brucellosis, syphilis, malaria

Answer 44

Toxoplasma Gondii- lymphocytosis with atypical lymphocytes is a hematologic hallmark of the disease

Answer 45

Mycobacterial tuberculosis

Answer 46

Know the written part

Answer 47

know the written

Answer 48

* Chronic lymphatic leukemia * Chronic myeloid leukemia * Acute lymphatic leukemia * Acute myeloid leukemia

Answer 49

Blood cells start as stem cells and then develop into specific types of white blood cells called lymphocytes * **Lymphocytes are made in the bone marrow** * become B cells (make antibodies) and T cells (attack and destroy bacteria, viruses, pathogens) * Can also be found in blood and lymph tissue

Answer 50

Lymph nodes are composed of **lymphocytes** * Found in clusters around the body * Become swollen or tender if you have an infection or malignancy

Answer 51

Bood cancers occur when healthy blood cells start dividing, growing or spreading abnormally

Answer 52

Leukemia * typically develops in lymphocytes but can also develop in myeloid cells * Leukemia cells can spread throughout the body but primarily affect the bone marrow and blood * Some are acute (grow quickly), others are chronic (grow slowly) Lymphoma * Develops in lymphocytes, however lymphoma usually affects lymph nodes and other organs instead of bone marrow * Separated into Hodgkin vs non-Hodgkin

Answer 53

* CLL is the most **common leukemia** in adult patients in the USA * Most often in people over age 55 * Lymphocytes are over 5000 cells/microL and are typically small mature looking with dense nuclei and compact chromatin, also called “soccer ball” cells * Numerous **smudge cells** on peripheral blood smear ## Footnote several genetic and chromosomal lesions paly a role in the malignant development of CLL B lymphocytes, in addition to antigens that could play a role in malignant cell selection.

Answer 54

* It is characterized by the uncontrolled proliferation of myeloid cells, particularly granulocytes (neutrophils, eosinophils, and basophils), in the bone marrow * The majority of cases of CML are associated with a specific genetic abnormality known as the **Philadelphia chromosome. This abnormality results from a translocation (exchange of genetic material) between chromosomes 9 and 22, leading to the formation of a fusion gene called BCR-ABL1.** The BCR-ABL1 fusion gene produces a mutant protein with abnormal tyrosine kinase activity, which plays a central role in the development and progression of CML.

Answer 55

* The diagnosis of CML is typically based on a combination of clinical evaluation, blood tests (complete blood count with differential), bone marrow biopsy, and genetic testing to detect the presence of the Philadelphia chromosome or BCR-ABL1 fusion gene. **The presence of the Philadelphia chromosome or BCR-ABL1 fusion gene is considered diagnostic of CML.** * Mainly affects adults

Answer 56

**Associated with increased lymphoblasts rather than more mature lymphocytes** **Acute lymphoblastic leukemia is classified based on the type of lymphocytes affected and the stage of maturation of the leukemic cells. The two main subtypes of ALL are**: * **B-cell ALL**: The majority of cases of ALL (about 85%) involve abnormal proliferation of immature B-cell lymphoblasts. * **T-cell ALL**: In this subtype, the leukemic cells are derived from abnormal proliferation of immature T-cell lymphoblasts.

Answer 57

Acute lymphoblastic leukemia is the most common type of leukemia in children, accounting for approximately 75-80% of pediatric leukemia cases. It is less common in adults but can still occur. The peak incidence of ALL occurs in children between 2 and 5 years of age

Answer 58

Diagnosis: CBC with differential, **bone marrow biopsy**, genetic testing for gene mutations associated with ALL, **flow cytometry (analyze the immunophenotype of the leukemic cells to distinguish between B-cell and T-cell ALL)**

Answer 59

* Affects blood cells called myeloid stem cells (myeloid can become one of 3 types of mature blood cells- RBC, WBC, platelets) * **Rapid abnormal proliferation and differentiation of these immature myeloid cells, called blasts, happens in the bone marrow** * In AML, the proliferation of immature myeloid blasts disrupts the normal production of these blood cell types so **healthy cells can’t do their job**

Answer 60

The incidence of AML increases with age, with the highest rates observed in individuals over 65 years old * BUT is second most common childhood leukemia after ALL Auer rod – a distinctive cytoplasmic inclusion found in AML * Abnormal structures formed from components of the cells cytoplasmic granules * Can also be seen in acute promyelocytic leukemia (that’s too far in detail!)

Answer 61

Several different types of non-Hodgkin lymphoma * Some start in B cells and others start in T cells * Is indolent or slow growing and slow spreading OR it can be aggressive

Answer 62

* Small mature looking lymphocytes and "smudge cells" in CLL and MBL * Atypical large lymphocytes are visible in EBV and other viral infections such as CMV or early HIV * Lymphocytes that are cleaved angulated, or have indented nuclei can be associated with pertussis or malignancies such as follicular lymphoma.

Answer 63

* "Hairy cells" with regular cytoplasmic projections are seen in hairy cell leukemia * Sezary Cells have cribriform nuclei with compact chromatin * "Villous" lymphocytes are seen in MZL * Large lymphocytes with multiple azurophilic granules are present in T-LGL * Lymphoblasts in ALL

Answer 64

* Flow cytometry * Fluorescence in situ hybridization (FISH)

Answer 65

* Powerful laboratory technique used to analyze and quantify various characteristics of cells, including their size, complexity, and protein expression, at the single-cell level * Cells of interest are collected from a biological sample, such as blood, bone marrow, tissue, or cell culture. These cells may be labeled with fluorescent dyes or antibodies targeting specific cell surface markers or intracellular proteins of interest

Answer 66

* The prepared cell sample is introduced into a flow cytometer, a sophisticated instrument equipped with lasers, optics, detectors, and fluidics systems. The flow cytometer can analyze thousands of cells per second as they pass through a narrow, focused stream of fluid, called the sheath fluid, within the instrument. * Laser Excitation: As each cell passes through the flow cytometer's laser beam, it is illuminated by one or more lasers of specific wavelengths. The laser light causes the fluorescent labels on the cells to emit light at characteristic wavelengths, which is then detected by photomultiplier tubes or other detectors.

Answer 67

Flow cytometry software enables researchers to identify different cell populations based on their unique characteristics, quantify the expression levels of specific proteins or markers, and perform complex analyses, such as cell cycle analysis or apoptosis assays.

Answer 68

Is a cytogenetic technique used to detect and analyze specific DNA sequences or chromosomal abnormalities in cells. Can visualize and quantify genetic alterations at the single-cell level.

Answer 69

* fluorescently labeled DNA or RNA probes are designed to specifically hybridize (bind) to complementary target sequences within the cell's genome. These probes can be designed to target specific genes, chromosomal regions, or DNA sequences of interest. * The sample containing cells of interest, typically obtained from blood, bone marrow, tissue biopsy, or cytological specimens, is first fixed onto a glass slide to preserve cellular morphology. The cells are then treated with chemicals or heat to denature (unwind) the DNA, allowing the probes to bind to their complementary target sequences * Excess unbound probes are washed away, leaving only the fluorescently labeled probes bound to their target sequences within the cells. The slide is then examined under a fluorescence microscope equipped with filters that can detect the fluorescence emitted by the bound probes. * The fluorescence signals emitted by the bound probes are visualized and captured using imaging software. The number, location, and intensity of the fluorescence signals provide information about the presence and characteristics of specific genetic abnormalities or chromosomal rearrangements within the cells *

Answer 70

* FISH can detect numerical abnormalities (aneuploidy) and structural abnormalities (translocations, deletions, duplications) in chromosomes associated with various genetic disorders, cancer, and developmental abnormalities. * **Prenatal Testing**: FISH can be used for prenatal diagnosis of chromosomal abnormalities, such as trisomy 21 (Down syndrome), trisomy 18 (Edwards syndrome), and trisomy 13 (Patau syndrome), by analyzing fetal cells obtained from amniotic fluid or chorionic villus sampling