Effect Of Methylprednisolone On Csf Igg Parameters Myelin Basic Protein And Anti
Published online by Cambridge University Press: 18 September 2015
- Kenneth G. Warren*
- Affiliation:Department of Medicine , Department of Laboratory Medicine and Rehabilitation Medicine of the University of Alberta, Edmonton
- Ingrid Catz
- Affiliation:Department of Medicine , Department of Laboratory Medicine and Rehabilitation Medicine of the University of Alberta, Edmonton
- Verona M. Jeffrey
- Affiliation:Department of Medicine , Department of Laboratory Medicine and Rehabilitation Medicine of the University of Alberta, Edmonton
- Dorothy J. Carroll
- Affiliation:Department of Medicine , Department of Laboratory Medicine and Rehabilitation Medicine of the University of Alberta, Edmonton
- *
- 9 – 101 Clinical Sciences Building ,
Statistical Analyses For Selection Of Proteins
Proteome Discoverer software was used to process the raw mass spectrometry files, identify the proteins and generate quantitative data which was further processed by three parallel approaches.
ANOVA-based
For each peptide, ANOVA was performed with the lmPerm R package to determine difference between groups. Afterwards, to determine which pairs of groups showed most differences, the Tukey’s HSD test was performed as post-hoc analysis.
Limma-based
Linear regression and analysis of variance were performed with the limma R package. The ratios of a specific protein between two compared groups were log2 transformed, normalized to the median, and the 3 replicates merged into one, and proteins were significant according to q-values . The resulting data were visualized in volcano plots and heatmaps using Perseus14.
Complementary analysis of the three replicates
Using the PD software, for each of the three sets the coefficient of variation CV of proteins within the set as well as the ratio of the mean abundance between the sets were calculated. A protein was selected for further analyses, if the ratio was larger than 1+2xCV . Subsequently, the PD software calculated a global ratio for a protein based on data from the three sets compared to healthy samples . Proteins were finally selected, if the protein expression was larger than 1+2xCV at least between two different conditions, and was consistently altered in a minimum of two of the three sets.
Cerebrospinal Fluid Protein Test
What is a cerebrospinal fluid protein test?
Cerebrospinal fluid is a clear bodily fluid that cushions and protects your brain and spinal cord. A CSF protein test involves taking a fluid sample from your spinal column using a needle. This procedure is known as a lumbar puncture or spinal tap.
The CSF protein test determines if theres too much or too little protein in your CSF. Test results that indicate your protein level is higher or lower than normal can help your doctor diagnose a range of conditions. Another use for a CSF protein test is to check the amount of pressure in your spinal fluid.
Don’t Miss: Best Protein Powders For Weight Loss
Does Ms Show Up In Blood Work
Blood tests cannot yet provide a definitive diagnosis of MS, although they can rule out other illnesses. Lyme disease is one of these additional disorders. Syphilis, tuberculosis, and HIV also may be detected through blood studies.
A blood test can tell whether you have antibodies against various diseases. These antigens are proteins that trigger the immune system to fight off infection. Your body makes antibodies against these proteins after being exposed to them. Having these antibodies does not necessarily mean that you have the disease associated with them, but it can help doctors diagnose problems or follow changes over time in those who have been diagnosed with such conditions.
Here are some examples of commonly tested antibodies:
Antibodies against cytomegalovirus : This virus is found in about half of all people. It usually causes no symptoms, but it can lead to serious health problems for certain groups of people, such as newborns, people with AIDS, and others who are immunocompromised. Testing for CMV antibodies helps doctors understand why you might be sick.
Antibodies against hepatitis B virus : HBV is the virus that causes hepatitis. Some people become chronic carriers of the virus and may never develop liver disease. Others may develop cirrhosis or cancer.
About Article Author
Silver Staining Of Agarose Gels
Agarose isoelectric focusing was carried out by using standard techniques. Briefly, isoelectric focusing of unconcentrated CSF was carried out at constant current using 1% zero-EEO agarose containing 10% ampholines, pH 3.5 to 10. At completion of the procedure, the gel was fixed in 60% methanol and 4% acetic acid, washed liberally in deionized water, dried using a hair dryer, and stained using silver as described below.
All reagents used for staining are described in Table 1. A 2% solution of potassium ferrocyanide was prepared in acetate buffer, pH 6, and a sufficient quantity was used to soak the dried gel. After 10 minutes, the gel was washed with copious amounts of deionized water, three times for 5 minutes each, for a total wash time of 15 minutes. Equal volumes of solutions A and B were mixed by a slow dropwise addition of solution B to solution A while avoiding the formation of a white precipitate. The gel was placed in this solution and the bands usually appeared within five minutes the development was complete in 15 minutes. After completion of the development, the gels were washed in 1% acetic acid for 20 minutes and dried.
Recommended Reading: Protein Rich Food For Veg
Flow Cytometry On Csf Cells
Fresh CSF was immediately spun down at 200 g for 10 min, the supernatant removed and the pellet resuspended in an appropriate volume of phosphate-buffered saline 5% FCS, depending on the white cell count of individual samples. A minimum number of 4000 cells was required for each staining combination, and a volume of 30 l was used for each staining. The staining was performed as described for whole blood cells . No lysing step was applied to CSF cells because no red blood cells were contained in these specimens. Cells were then washed once with PBS 5% FCS and analysed as described above. Because of the small cell number obtained from control donors and some multiple sclerosis patients, staining combinations and were not performed in every patient.
Are There Any Risks To Having A Csf Protein Test
ââSerious risks are rare with the procedure, but you may have more minor side effects.
These can include:
- âHeadaches called âpost-lumbar headaches,â which can last for a few hours to a week
- âPain or tenderness where the needle went in
- âSlight bleeding at the wound site
Rarely, you may get more serious side effects like infections, inflammation, or severe bleeding. Speak to your doctor or head to an emergency room immediately if you feel unwell.
Recommended Reading: Which Protein Powder Is Best For Muscle Gain
What Is The Test
To detect OCB levels, a doctor may order a CSF oligoclonal banding test, which requires a sample of the CSF. The doctor will collect the sample by performing a lumbar puncture, or spinal tap.
People who receive a diagnosis of MS typically have positive test results, indicating that OCB is present in the CSF.
The test alone is not enough to diagnose MS. A doctor must also find:
- at least two areas of damage to, or lesions in, the brain or spinal cord
- evidence that the damage occurred at different times
- evidence that another disease has not caused the damage to the CNS
Cerebrospinal Fluid Biomarkers Of Disease Activity
The assessment of disease activity is useful to guide therapeutic interventions. At present, this assessment is based on clinical relapses, disability progression, and the appearance of new lesions in magnetic MRI.99. Rotstein DL, Healy BC, Malik MT, Chitnis T, Weiner HL. Evaluation of no evidence of disease activity in a 7-year longitudinal multiple sclerosis cohort. JAMA Neurol. 2015 72:152-8. Reliable biomarkers reflecting subclinical disease are still lacking.
You May Like: Diamond High Protein Dog Food
Conditions Associated With Changes In The Biochemical Composition Of Csf
Conditions associated with changes in glucose concentration of CSF include the following:
- Bacterial, fungal, and tuberculous meningitis
- Primary or metastatic meningeal malignancy
Note that normal concentration of glucose in CSF samples is 45-80 mg/dL or 60-80% of that in the plasma . Absolute decreased CSF glucose level and especially decreased CSF glucose level in relation with serum are usually associated with bacterial or fungal meningitis. However, in patient with a normal CSF glucose concentration but with increased number of WBC, viral meningitis should be suspected. For accurate interpretation of CSF glucose concentration, serum glucose should be evaluated in serum samples collected about 2 hours prior to spinal tapping and all specimens should be tested immediately to avoid glycolysis.
Conditions associated with elevated CSF lactate include any condition associated with decreased blood flow or hypoxia , such as the following:
- Mycoplasma meningitis
Evaluation of lactic acid concentration in CSF is useful for the diagnosis and management of different types of meningitis. Generally, the following guidelines can be applied:
- CSF lactate > 35 mg/dL is seen in patients with bacterial meningitis
- CSF lactate 25-35 mg/dL is seen in patients with tubercular and fungal meningitis
- CSF lactate < 25 mg/dL is seen in patients with viral meningitis
Conditions associated with elevated CSF LDH include the following:
- Intracranial hemorrhage
- Bacterial meningitis
How Is A Spinal Tap Performed
A spinal tap is a procedure used to collect a sample of CSF. This sample is then analyzed in a lab.4
The test involves inserting a needle into the spinal canal. You will either lie on your side with your knees drawn up to your chest or sit up and lean over with your back arched. These positions help open up the spinal canal and make it easier for the doctor to insert the needle.4
The doctor will insert the needle between the vertebrae in the lower back. Nerves that branch off of the spinal cord are located in this area.4
A spinal tap is performed using local anesthesia. This numbs the area and helps to decrease discomfort. Local anesthesia causes brief stinging and burning when injected but then numbs the area. It takes about 30 minutes to complete the procedure.4
Don’t Miss: High Protein Snacks For Muscle Gain
What Do The Results Mean
A CSF analysis may include a variety of different tests on your sample. So, the measurements on your test results will depend on which tests were done. Your provider can explain what your results mean.
In general, your CSF analysis results may show that you have an infection, an autoimmune disorder, such as multiple sclerosis , or another disease or injury of the brain or spinal cord. Your provider will likely order more tests to confirm your diagnosis.
Learn more about laboratory tests, reference ranges, and understanding results.
What Abnormal Results Mean
An abnormal protein level in the CSF suggests a problem in the central nervous system.
Increased protein level may be a sign of a tumor, bleeding, nerve inflammation, or injury. A blockage in the flow of spinal fluid can cause the rapid buildup of protein in the lower spinal area.
A decrease in protein level can mean your body is rapidly producing spinal fluid or you have a more generalized protein deficiency.
Recommended Reading: Good Protein Powder For Muscle Gain
Summary Of Chemistry Evaluation Of Csf In Different Clinical Conditions
Chemistry evaluation of CSF in different clinical conditions is summarized below.
Table. Changes in Analytes With Various CNS Disease
|
Disease |
Tube 1 is generally used for Chemistry and some hematology testing.
In the case of a traumatic tap for CSF collection, the first collection tube will contain CSF with a high number of RBC and high concentration of hemoglobin. Presence of these compounds can lead to inaccurate measurements for some chemistry analytes. Therefore specific corrections must be considered. can be used for chemistry testing.
Similarly, if cell count is evaluated using first tube CSF collection from a traumatic tap, the RBC count will be significantly elevated resulting in misleading results. Therefore, in these circumstances, tube 4 should be used instead.
Tube 2 is generally used for microbiology testing. It can also be used for molecular testing, and viral or serological testing.
Tube 3 is generally used for cytology.
Tube 4 is generally used for hematology and flow cytometry.
Additional tubes can be collected and used for more esoteric testing .
Lumbar puncture is contraindicated in patients with neurological signs indicating increased intracranial pressure .
Csf Findings In Other Inflammatory Demyelinating Diseases
At first onset some symptoms are similar between MS and other inflammatory demyelinating diseases, particularly neuromyelitis optica spectrum diseases. These syndromes can be diagnosed by IgG antibodies against AQP4 or MOG . In general CSF work-up there is a distinct feature, which is a lack of CSF OCB in NMO spectrum diseases in 8090% of patients . Total leukocyte counts in NMO spectrum disorders are similar to MS with pleocytosis being found in around 50% of patients, exceeding rarely 100 cells per uL . However, on differential cell counts granulocytes occur somewhat more frequently in NMO spectrum disorders compared to MS .
In MOG-IgG antibody associated syndromes the frequency of OCB of 13% is similarly low as in NMO spectrum disorders . It seems however, that CSF pleocytosis occurs more frequently, i.e., in almost two thirds of patients with a relatively high proportion of neutrophils making up 22% of all leukocytes . Also, an elevated albumin quotient can be found in roughly one third of patients with MOG IgG antibodies, particularly if spinal symptoms occur.
Altogether, the main distinctive feature between these syndromes and MS is the frequency of OCB, whereas general CSF changes differ slightly but do not provide compelling evidence for or against one of the entities.
Don’t Miss: What Has Protein Besides Meat
Combining The Data Of All Individuals Within Each Group Across Cohort 1 And 2
The proteome data from the two cohorts were combined to perform confidence interval analyses within each group across the cohorts . This revealed a highly consistent pattern of variation as the analysis was performed by simply merging the data for the groups from the two independent cohorts. Thus, the proteins identified by analysis of the cohorts discriminated MS/CIS patients from controls irrespectively of intrathecal inflammation and irrespectively of the stage of the disease as CIS or MS. As there was more power in the statistical analysis of the combined cohorts due to the larger number of individuals, the number of significant proteins is higher for the combined data sets across the two cohorts than for similar analyses performed within the cohorts .
Comparison Of Csf And Blood Leucocyte Populations From Multiple Sclerosis Patients And Control Donors
CSF and PB from 60 multiple sclerosis patients with clinically definite multiple sclerosis or laboratory-supported definite multiple sclerosis were analysed. Multiple sclerosis patients were compared with a group of 21 patients with NIND. The initial comparison included levels of protein, albumin, IgG, IgA and IgM in serum and CSF. In addition, we determined the phenotype of the cellular compartments using monoclonal antibodies specific for CD45 , CD3 , CD19 , CD138 , CD16/CD56 , CD14 , CD4 , CD8 , TCR-/ and TCR-/. These mAbs enabled determination of the absolute number of each cell type and the relative percentage of all subpopulations from the pool of leucocytes in PB and CSF.
First, we compared leucocyte subsets in PB between multiple sclerosis patients and NIND patients. Although intra-individual variations were noticed for all leucocyte subsets, no significant differences were observed between multiple sclerosis and NIND patients in PB . In contrast, highly significant differences were observed for immunoglobulin levels and leucocyte subsets in CSF between multiple sclerosis and NIND patients. As expected, the IgG levels and the absolute white cell count were increased in CSF from multiple sclerosis patients . We found a higher proportion of B cells and plasma cells in the CSF of multiple sclerosis patients as reported previously . The percentage of monocytes, NK cells and NK-like T cells was decreased in multiple sclerosis patients compared with NIND patients .
Also Check: Cookies And Cream Protein Shakes
How The Test Is Performed
A sample of CSF is needed . Alumbar puncture is the most common way to collect this sample. Rarely, other methods are used for collecting CSF such as:
- Cisternal puncture
- Ventricular puncture
- Removal of CSF from a tube that is already in the CSF, such as a shunt or ventricular drain.
After the sample is taken, it is sent to a lab for evaluation.
Is Hsp Similar To Ms
HSP has numerous clinical similarities with the progressive phase of Multiple Sclerosis , a chronic neurodegenerative disease marked by inflammation, demyelination, and axonal loss in multiple parts of the central nervous system . Similar symptoms include muscle weakness, pain, tremors, and problems with coordination. However there are also differences between HSP and MS. For example, people with HSP do not usually experience any problems with vision or balance, for example.
HSP is not just one disease, but rather a group of diseases caused by mutations in different genes that lead to a common symptom pattern. The three main types of HSP are known as “classic” or “genetic” HSP, non-classical HSP, and inclusion body myopathy . Genetic HSP occurs when one of the four known gene mutations is present. These mutations affect proteins that help form normal stress fibers and abnormal stress fibers called hyperstructures. Non-classical HSP includes all other types of HSP except classic HSP. These other types include sporadic HSP, which is not inherited acquired HSP, which can be caused by physical trauma to the skin or muscles and refractory HSP, which does not respond to treatment. IBM is a rare type of muscular dystrophy that often starts in adulthood and causes severe muscle damage due to the formation of protein inclusions inside muscle cells.
Don’t Miss: Protein Shakes Without Protein Powder