Hodgkin lymphoma (HL) is characterized by constitutive activation of several signaling pathways and expression of a distinct set of transcription factors. This characteristic phenotype is partly caused by gene mutations as determined by targeted and whole genome sequencing approaches on cell lines and microdissected HRS cells. Here we studied CD58 and MYB, 2 genes we previously identified by whole exome sequencing (WES) in HL cell lines. CD58 gene mutations observed in 3 HL cell lines were confirmed by Sanger sequencing at the DNA and RNA level. CD58 protein expression as determined by flow, western blot and IHC was absent in all 3 HL cell lines with mutations and present in 4 HL cell lines with wild type CD58. In primary tissue samples, loss of CD58 expression was observed in 11% of the patients. Lack of CD58 was specifically observed in patients who developed a relapse. This suggests that loss of CD58 is a potential immune escape mechanism of HRS tumor cells, especially in clinically aggressive disease. Frame shift mutations resulting in a truncated MYB protein were found in L428 and SUPHD1 cells. The mutation in L428 causes complete loss of most of the C-terminal domain whereas the mutation in SUPHD1 results in a MYB protein with a partial loss of the C-terminal domain. We confirmed the mutations at the DNA and RNA level and confirmed presence of a truncated protein by Western blot with C-terminal and N-terminal domain specific antibodies. RT-PCR with primers specific for each of the alternatively spliced exons, indicated that these were not included in the MYB transcript in any of the HL cell lines. In tissue samples, staining of MYB in HL cases revealed no staining with both antibodies in the vast majority of the cases, indicating complete loss of the MYB protein. Inhibition of MYB by shRNA constructs induced negative effects on cell growth in HL cell lines with wild type MYB (L540 & KMH2) and no or limited effects in HL cell lines with truncated MYB (L428 & SUPHD1). Microarray analyses of L540 & KMH2 cells treated with non-targeting and MYB shRNA constructs revealed 63 consistently MYB induced and repressed genes. In conclusion, we show that loss of CD58 expression is common in HL cell lines and restricted to the HRS cells of tissue samples of relapsed HL. For MYB, strong oncogenic effects were observed especially for the HL cell lines with wild type MYB.