IMMUNOLOGY ASSIGNMENT

IMMUNOPHENOTYPING

This technique finds application in fundamental research and aiding disease detection, like
certain forms of leukemia and lymphoma. Additionally, immunophenotyping can be employed
to categorize cells into distinct groups based on the surface markers they possess.

Immunophenotyping detects the presence or absence of white blood cell (WBC) antigens. These
antigens are protein structures found on the surface or interior of WBCs.

Normal white blood cells (WBCs) display typical antigen groupings, while specific leukemias
and lymphomas exhibit distinctive but abnormal antigen patterns. As a result,
immunophenotyping proves valuable in diagnosing and categorizing these blood cell cancers
based on the unique presence or absence of certain antigens.

Flow cytometry immunophenotyping may be useful in helping to diagnose, classify, treat, and
determine prognosis of these blood cancers.

IMMUNOPHENOTYPING OF LEUKAEMIA AND LYMPHOMA

Leukemias and lymphomas arise from an aberrant white blood cell that undergoes uncontrolled
division, leading to the proliferation of multiple identical copies of itself, known as clones.

LEUKAEMIA

Leukemias result from the abnormal replication of specific types of white blood cells, known as
lymphocytes or myeloid (granular) white blood cells. This uncontrolled cloning process gives
rise to monoclonal lymphocytic or myeloid leukemias.

LYMPHOMA

Lymphomas develop due to the abnormal proliferation of lymphocytes, resulting in a

monoclonal population and leading to cancer affecting the lymphatic system.

The two main kinds of lymphoma are—

• Hodgkin lymphoma; exhibits a methodical pattern of spreading, moving systematically

from one group of lymph nodes to another..
 • Non-Hodgkin lymphoma, in contrast, spreads throughout the lymphatic system in a non-
orderly or unpredictable manner.

Abnormal
Abnormal Inhibit WBCs, RBCs &
cells increases
cells grow platelets production
in lymph node

Don't fight Abnormal cells size of lymph node

infections increases in BM increases

Don't die
Accumulation occur
at normal in BM, Lymph node Lymphpma
rate

FLOW CYTOMETRY IMMUNOPHENOTYPING

Flow cytometry immunophenotyping may be performed on

• blood,
• bone marrow,
• or other samples to provide this additional information.

Immunophenotyping is capable of detecting both normal and abnormal cells, recognizing

specific marker patterns associated with certain types of leukemia and lymphoma. Moreover,
the results can provide insights into the cancer's aggressiveness and its likelihood of
responding to particular treatments.

Flow cytometry immunophenotyping is the method employed to diagnose and categorize

blood cell cancers such as leukemias and lymphomas. An increased lymphocyte count,
presence of immature blood cells, or abnormal cell counts in a complete blood count (CBC)
and white blood cell (WBC) differential may prompt further investigation.
Flow cytometry immunophenotyping may also be used:

To predict how aggressive the cancer will be

• To predict whether the cancer will respond to certain treatment

To help determine whether treatment of leukemia or

lymphoma has been successful

 In determining the persistence of the disease following treatment (residual

disease) or its recurrence after a successful treatment (recurrent disease).


Flow cytometry is carried out through the following steps:

• Processing a blood sample, bone marrow (BM), tissues, or other fluid to remove red blood
cells.

• Introducing antibodies labeled with fluorescent markers.

• These antibodies attach to specific antigens present on white blood cells.The WBCs are then
drawn up into a single-cell fluid stream under pressure past multiple lasers and detectors and
each cell is analysed individually.

When it is necessary to used this test?

Flow cytometry immunophenotyping might be requested when there is a rise in lymphocyte count (or
occasionally an elevation in other white blood cell types, WBCs), presence of anemia, reduced platelet
count, or the presence of immature WBCs not typically observed in the blood. These indicators could be
the initial signs of a potential blood cell cancer.
 SIGNS AND SYMPTOMS
Signs and symptoms of blood cell cancer include:

weight shortness
weakness pale skin
loss of breath

bone and enlarged

fever headache
joint pain organs

night
vomiting
sweats

RESULT:

The outcome of flow cytometry immunophenotyping assists in characterizing the cells by

identifying the markers present on them. Normal cells exhibit a specific pattern of antigens
corresponding to their type and maturity.

Your immunophenotyping results are then compared to the antigen patterns of both normal cells
and abnormal cells associated with leukemias and lymphomas.

 In an older child or adult's blood, mature B cells are usually present, but circulating immature
B cells are not typical. Both mature and immature B cells show positivity for the CD19
marker. Mature B cells are positive for CD20, indicating maturity, while CD34 is not
expressed. CD34 serves as a marker for immaturity, and normally, CD20 and CD34 are not co-
expressed.

 If an adult displays a small number of mature B cells along with a large number of immature
B cells that are positive for CD19 (a B-cell marker) and also positive for both CD34 and CD20
(indicating immaturity and abnormality), then the individual is diagnosed with an immature
B-cell leukemia known as B-lymphoblastic leukemia.

 Interestingly, certain additional antigens detected during immunophenotyping may indicate a

specific genetic subtype of B-lymphoblastic leukemia, potentially influencing the prognosis.
  Furthermore, there are newer and advanced treatment options available, such as CAR-
T therapy, bispecific T-cell engagers, and monoclonal antibodies, which selectively
target molecules like CD19 or CD20. These innovative therapies may result in
reduced side effects compared to conventional chemotherapy, and they are often used
in combination with traditional chemotherapy for enhanced efficacy.

ABNORMAL IMMUNOPHENOTYPE

Abnormal immunophenotype profiles are usually present in:

 Acute myeloid leukemia

 Acute lymphoblastic lymphoma (ALL)
 Chronic lymphocytic leukemia (CLL)
 B-cell and T-cell non-Hodgkin Lymphomas
 Multiple myeloma