Gliomas are tumors of the brain parenchyma that are classified histologically based on their resemblance to different types of glial cells. In Iraq, as in many countries, gliomas pose a significant health challenge, the spread of glioma disease refers to its incidence and prevalence within the population progressively. Material and methods: Fixed paraffin-embedded Glioma brain tumors of 64 samples, thirty-four as patients (21 male and 13female) and thirteen(benign) as control (10 male and 20 female) were collected from the Neurological Hospital in Baghdad, samples date from December 2022 to July 2023. Diagnosis and grading of Glioma (high grade) in the study were done by immunohistochemistry technique by Nestin marker to investigate its expression in samples through a scoring system where it was, 0 negative, 1 light, 2 moderate, and 3 strong. Results: The result showed non-significant P≤0.01 between male and female inpatient p-value (0.277), with mean (3.29±0.56, 3.69±0.13, respectively), noticed a high significant P≤0.01 between patient and control p-value (0.0001) in both sex, it also showed highly significant difference P≤0.01 in the percentage of patients distribution according to scores, and the highest percentage for male and female was at score 2 and 3, in percentages for male (42.86) and female (30.77, 69.23, respectively). Conclusion: The results confirm that Nestin is associated with high-grade Glioma.